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Protective effects of anti-ricin A-chain RNA aptamer against ricin toxicity

World Journal of Gastroenterology 2008年第41期14卷 6360-6365页

作者： Shaoan Fan Feng Wu Frank Martiniuk Martha L Hale andrew d ellington Kam-Meng Tchou-Wong department of Environmental Medicine New York University School of Medicine 57 Old Forge Road Tuxedo New York 10987 United States department of Medicine New York University School of Medicine 550 First Avenue New York 10016 United States Integrated Toxicology division U.S. Army Medical Research Institute of Infectious Diseases (USAMRIID) Fort Detrick Maryland 21702 United States department of Chemistry and Biochemistry University of Texas at Austin Austin Texas 78712 United States

AIM： To investigate the therapeutic potential of an RNA ligand （aptamer） specific for the catalytic ricin A-chain （RTA）, the protective effects of a 31-nucleo- tide RNA aptamer （31RA）, which formed a high affinity complex with RTA, against ricin-induced toxicity in cell- based luciferase translation and cell cytotoxicity assays were evaluated. METHOdS： To test the therapeutic potential of anti- RTA aptamers in Chinese hamster ovary （CliO） AA8 cells stably transfected with a tetracycline regulatable promoter, ricin ribotoxicity was measured us- ing luciferase and ricin-induced cytotoxicity was ascertained by MTS cell proliferation assay with tet- razolium compound [3-（4,5-dimethylthiazol-2-yl）- 5-（3-carboxymethoxyphenyl）-2-（4-sulfophenyl）-2H- tetrazolium]. RESULTS： Inhibition of protein synthesis by ricin in CliO AA8 cells resulted in diminished luciferase activity and treatment with polyclonal antibody against degly- cosylated RTA （dgA） neutralized the inhibitory effects of ricin on luciferase activity and protected against ricin-induced cytotoxicity as measured by MTS assay. The 31RA anti-RTA aptamer inhibited the translation of luciferase mRNA in cell-free reticulocyte translation assay. 31RA aptamer also partially neutralized the inhibitory effects of ricin on luciferase activity and partially protected against ricin-induced cytotoxicity in CliO AA8 cells. CONCLUSION： We have shown that anti-RTA RNA aptamer can protect against ricin ribotoxicity in cell- based luciferase and cell cytotoxicity assays. Hence, RNA aptamer that inhibits RTA enzymatic activity represents a novel class of nucleic acid inhibitor that has the potential to be developed as a therapeutic agent for the treatment of ricin intoxication.

关键词： Ricin inhibitor RNA aptamer Luciferase assay

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Phosphorothioated primers lead to loop-mediated isothermal amplification at low temperatures

Phosphorothioated primers lead to loop-mediated isothermal a...

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中国化学会第十三届全国分析化学年会

作者： Sheng Cai Cheulhee Jung Sanchita Bhadra andrew d.ellington Laboratory of Pharmaceutical Analysis and drug Metabolism Zhejiang Province Key Laboratory of Anti-Cancer Drug Research Zhejiang University Institute for Cellular and Molecular Biology Department of Molecular Biosciences University of Texas at Austin

Loop-mediated isothermal amplification（LAMP） is an extremely powerful tool for the detection of nucleic acids with high sensitivity and specificity. LAMP shows best performance at around 65 °C, which is relatively high to be easily applied to point-of-care-testing（POCT）. Here, we developed a phosphorothioated LAMP（PS-LAMP） working at low temperature by providing a more efficient amplification path. PS-modifications enabled efficient self-folding at the termini to generate more loops where intact inner primers bind and extend. By optimizing several factors such as urea, Bst 2.0 dNA polymerase, single-stranded dNA binding protein（SSB） and MgSO4, comparable sensitivity and selectivity with a regular LAMP（R-LAMP） at 65 °C were achieved with a PS-LAMP at 40 °C. As the novel PS-LAMP system is performed at around physiological temperature, it has a great potential in applications to hand-held or POCT diagnostic devices.

关键词： Phosphorothioated primers LAMP POCT

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Male and female meadow voles Microtus pennsylvanicus respond differently to scent marks from the top- middle-, and bottom-scent donors of an over-mark

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Current Zoology 2011年第4期57卷 441-448页

作者： Michael H. FERKIN Nicholas J. HOBBS Benjamin d. FERKIN Adam C.FERKIN daniel A. FERKIN department of Biological Sciences The University of Memphis Ellington Hall Memphis TN 38152 USA

Previous studies have shown that individuals responded preferentially to the mark of the top-scent donor relative to that of the bottom-scent donor of an over-mark. However, terrestrial mammals are likely to encounter over-marks consisting of the scent marks of more than two same-sex conspecifics in the intersections of runways, near the nests of sexually receptive female conspecifics, and inside and along the borders of the territories of conspecifics. We determined how meadow voles, Microtus pennsylvanicus, respond to the marks of the top-, middle-, and bottom-scent donors of an over-mark. We tested the hypothesis that voles exposed to an over-mark will respond preferentially to the scent marks that were deposited more recently, the scent marks that were on top or near the top of the over-mark, compared to the scent marks that were deposited earlier or near the bot- tom of the over-mark. Voles spent more time investigating the mark of the top-scent donor than that of the either the middle- or bottom-scent donor. However, males but not female voles spent more time investigating the middle-scent mark than the bot- tom-scent mark. We also tested the hypothesis that voles evaluate and respond to over-marks differently from single scent marks. Voles spent more time investigating the marks of the top-, middle-, and bottom-scent donors compared to scent marks that were not part of the over-mark. Voles can distinguish among the overlapping scent marks of three scent donors and sex differences exist in the values they appear to attach to each of these scent marks [Current Zoology 57 （4）： 441-448, 2011 ].

关键词： Over-marks Voles Scent marks Olfactory communication

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