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101.
Cooley M Carychao D Crawford-Miksza L Jay MT Myers C Rose C Keys C Farrar J Mandrell RE 《PloS one》2007,2(11):e1159
Fresh vegetables have become associated with outbreaks caused by Escherichia coli O157:H7 (EcO157). Between 1995-2006, 22 produce outbreaks were documented in the United States, with nearly half traced to lettuce or spinach grown in California. Outbreaks between 2002 and 2006 induced investigations of possible sources of pre-harvest contamination on implicated farms in the Salinas and San Juan valleys of California, and a survey of the Salinas watershed. EcO157 was isolated at least once from 15 of 22 different watershed sites over a 19 month period. The incidence of EcO157 increased significantly when heavy rain caused an increased flow rate in the rivers. Approximately 1000 EcO157 isolates obtained from cultures of>100 individual samples were typed using Multi-Locus Variable-number-tandem-repeat Analysis (MLVA) to assist in identifying potential fate and transport of EcO157 in this region. A subset of these environmental isolates were typed by Pulse Field Gel Electrophoresis (PFGE) in order to make comparisons with human clinical isolates associated with outbreak and sporadic illness. Recurrence of identical and closely related EcO157 strains from specific locations in the Salinas and San Juan valleys suggests that transport of the pathogen is usually restricted. In a preliminary study, EcO157 was detected in water at multiple locations in a low-flow creek only within 135 meters of a point source. However, possible transport up to 32 km was detected during periods of higher water flow associated with flooding. During the 2006 baby spinach outbreak investigation, transport was also detected where water was unlikely to be involved. These results indicate that contamination of the environment is a dynamic process involving multiple sources and methods of transport. Intensive studies of the sources, incidence, fate and transport of EcO157 near produce production are required to determine the mechanisms of pre-harvest contamination and potential risks for human illness. 相似文献
102.
Mark C. Ball Richard Pither Micheline Manseau Jeff Clark Stephen D. Petersen Steve Kingston Natasha Morrill Paul Wilson 《Conservation Genetics》2007,8(3):577-586
Faecal material has increasingly become an important non-invasive source of DNA for wildlife population genetics. However,
DNA from faecal sources can have issues associated with quantity (low-template and/or low target-to-total DNA ratio) and quality (degradation and/or low DNA-to-inhibitor ratio). A number of studies utilizing faecal material assume
and compensate for the above properties with minimal characterization of quantity or quality of target DNA, which can unnecessarily increase the risk of downstream technical problems. Here, we present a protocol which quantifies
faecal DNA using a two step approach: (1) estimating total DNA concentration using a PicogreenTM fluorescence assay and (2) estimating target nuclear DNA concentration by comparing amplification products of field samples at suspected concentrations to those of control
DNA at known concentrations. We applied this protocol to faecal material collected in the field from two species: woodland
caribou (Rangifer tarandus) and swift fox (Vulpes velox). Total DNA estimates ranged from 6.5 ng/μl to 28.6 ng/μl (X = 16.2 ng/μl) for the caribou extracts and 1.0–26.1 ng/μl (X = 7.5 ng/μl) for the swift fox extracts. Our results showed high concordance between total and target DNA estimates from woodland caribou faecal extracts, with only 10% of the samples showing relatively lower target-to-total DNA ratios. In contrast, DNA extracts from swift fox scat exhibited low target DNA yields, with only 38% (19 of 50) of the samples showing comparative target DNA amplification of at least 0.1 ng. With this information, we were able to estimate the amount of target DNA entered into PCR amplifications, and identify samples having target DNA below a lower threshold of 0.2 ng and requiring modification to genotyping protocols such as multiple tube amplification.
Our results here also show that this approach can easily be adapted to other species where faeces are the primary source of
DNA template. 相似文献
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Leigh‐Ann Woolley Hayley M. Geyle Brett P. Murphy Sarah M. Legge Russell Palmer Christopher R. Dickman John Augusteyn Sarah Comer Tim S. Doherty Charlie Eager Glenn Edwards Dan K.P. Harley Ian Leiper Peter J. McDonald Hugh W. McGregor Katherine E. Moseby Cecilia Myers John L. Read Joanna Riley Danielle Stokeld Jeff M. Turpin John C.Z. Woinarski 《Mammal Review》2019,49(4):354-368
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106.
Timothy F. Czajka David J. Vance Simon Davis Michael J. Rudolph Nicholas J. Mantis 《The Journal of biological chemistry》2022,298(4)
During ricin intoxication in mammalian cells, ricin''s enzymatic (RTA) and binding (RTB) subunits disassociate in the endoplasmic reticulum. RTA is then translocated into the cytoplasm where, by virtue of its ability to depurinate a conserved residue within the sarcin–ricin loop (SRL) of 28S rRNA, it functions as a ribosome-inactivating protein. It has been proposed that recruitment of RTA to the SRL is facilitated by ribosomal P-stalk proteins, whose C-terminal domains interact with a cavity on RTA normally masked by RTB; however, evidence that this interaction is critical for RTA activity within cells is lacking. Here, we characterized a collection of single-domain antibodies (VHHs) whose epitopes overlap with the P-stalk binding pocket on RTA. The crystal structures of three such VHHs (V9E1, V9F9, and V9B2) in complex with RTA revealed not only occlusion of the ribosomal P-stalk binding pocket but also structural mimicry of C-terminal domain peptides by complementarity-determining region 3. In vitro assays confirmed that these VHHs block RTA–P-stalk peptide interactions and protect ribosomes from depurination. Moreover, when expressed as “intrabodies,” these VHHs rendered cells resistant to ricin intoxication. One VHH (V9F6), whose epitope was structurally determined to be immediately adjacent to the P-stalk binding pocket, was unable to neutralize ricin within cells or protect ribosomes from RTA in vitro. These findings are consistent with the recruitment of RTA to the SRL by ribosomal P-stalk proteins as a requisite event in ricin-induced ribosome inactivation. 相似文献
107.
Adrienne H K Roeder Marisa S Otegui Ram Dixit Charles T Anderson Christine Faulkner Yan Zhang Maria J Harrison Charlotte Kirchhelle Gohta Goshima Jeremy E Coate Jeff J Doyle Olivier Hamant Keiko Sugimoto Liam Dolan Heather Meyer David W Ehrhardt Arezki Boudaoud Carlos Messina 《The Plant cell》2022,34(1):72
As scientists, we are at least as excited about the open questions—the things we do not know—as the discoveries. Here, we asked 15 experts to describe the most compelling open questions in plant cell biology. These are their questions: How are organelle identity, domains, and boundaries maintained under the continuous flux of vesicle trafficking and membrane remodeling? Is the plant cortical microtubule cytoskeleton a mechanosensory apparatus? How are the cellular pathways of cell wall synthesis, assembly, modification, and integrity sensing linked in plants? Why do plasmodesmata open and close? Is there retrograde signaling from vacuoles to the nucleus? How do root cells accommodate fungal endosymbionts? What is the role of cell edges in plant morphogenesis? How is the cell division site determined? What are the emergent effects of polyploidy on the biology of the cell, and how are any such “rules” conditioned by cell type? Can mechanical forces trigger new cell fates in plants? How does a single differentiated somatic cell reprogram and gain pluripotency? How does polarity develop de-novo in isolated plant cells? What is the spectrum of cellular functions for membraneless organelles and intrinsically disordered proteins? How do plants deal with internal noise? How does order emerge in cells and propagate to organs and organisms from complex dynamical processes? We hope you find the discussions of these questions thought provoking and inspiring.We asked 15 experts to address what they consider to be the most compelling open questions in plant cell biology and these are their questions. 相似文献
108.
Antonio Garrido Montalban Erik Boman Chau-Dung Chang Susana Conde Ceide Russell Dahl David Dalesandro Nancy G.J. Delaet Eric Erb Justin T. Ernst Andrew Gibbs Jeffrey Kahl Linda Kessler Jeff Kucharski Christopher Lum Jan Lundström Stephen Miller Hiroshi Nakanishi Edward Roberts Eddine Saiah Robert Sullivan Christopher J. Larson 《Bioorganic & medicinal chemistry letters》2010,20(16):4819-4824
We have optimized a novel series of potent p38 MAP kinase inhibitors based on an α-ketoamide scaffold through structure based design that due to their extended molecular architecture bind, in addition to the ATP site, to an allosteric pocket. In vitro ADME, in vivo PK and efficacy studies show these compounds to have drug-like characteristics and have resulted in the nomination of a development candidate which is currently in phase II clinical trials for the oral treatment of inflammatory conditions. 相似文献
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