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Atlanta Margarita + Taco Festival Group

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Richard Savin
Richard Savin

La Cr Offsite 2013 __TOP__

The February 2013 North American blizzard, also known as Winter Storm Nemo[5][6] and the Blizzard of 2013,[7] was a powerful blizzard that developed from the combination of two areas of low pressure,[8] primarily affecting the Northeastern United States and parts of Canada, causing heavy snowfall and hurricane-force winds. The storm crossed the Atlantic Ocean, affecting Ireland and the United Kingdom.[9] The nor'easter's effects in the United States received a Category 3 rank on the Northeast Snowfall Impact Scale, classifying it as a "Major" Winter Storm.

La Cr Offsite 2013

The first low-pressure system, originating from the Northern Plains of the United States, produced moderate amounts of snow across the Great Lakes region of the U.S. and Canada.[10] The second low, originating across the state of Texas, produced heavy rains and flooding across much of the Southeast and Mid-Atlantic parts of the U.S. As the two systems merged off the Northeast coast on February 8, 2013, they produced heavy snowfall over a large region from North Jersey and inland from New York City through eastern New England up to coastal Maine and inland to Ontario.

Atmospheric conditions leading up to the formation of the February 2013 nor'easter were rather anomalous and were conducive for cyclogenesis. Such conditions included the presence of a ridge over the North Atlantic and the strengthening of a trough in California.[25] The impactful nature of the eventual winter storm event was first predicted in National Centers for Environmental Prediction (NCEP) and Hydrometeorological Prediction Center (HPC) forecasts at 1200 UTC on February 6,[nb 1] when the accumulation of at least 4 in (100 mm) of snow in the ensuing hours was considered a moderate probability in parts of Wisconsin and Michigan.[26] Fueled by energy originating from the Gulf of Alaska and carried by the polar jet stream, a low-pressure area formed as anticipated in Montana, at 1200 UTC the next day.[25] The cyclone tracked in a general east-northeastward direction throughout the day,[27] reaching central Indiana by early on February 8.[28]

By late on February 7, 2013, winter storm warnings and winter weather advisories were issued for the northeastern United States, from the Upper Midwest to New England. There was also a blizzard warning for the New York metropolitan area, all of Connecticut, all of Rhode Island, and eastern Massachusetts, as well as southeast New Hampshire and coastal Maine.[32] On February 8, blizzard warnings were expanded to include inland portions of southeast New Hampshire, and inland portions of Maine's coastal counties.[51] By February 8, storm warnings and hurricane force wind warnings were in effect for the New England and Mid Atlantic waters, in addition to coastal flood warnings.[52]

The Pilgrim Nuclear Power Plant lost power again on February 10, after an offsite outage had caused a shutdown on February 8. A spokeswoman for Entergy Corp., the Louisiana company that owns Pilgrim said, "There's no worker or public safety concern," she said. "We're troubleshooting the cause."[92]

The Weather Channel dubbed the storm "Winter Storm Nemo",[11][5][115][116][6][117] in keeping with a list of names they have given to some winter storms since 2012.[118] Hartford, Connecticut, CBS affiliate WFSB named the storm "Blizzard Charlotte", in keeping with a long-standing station tradition of naming major winter storms affecting Connecticut dating back to the early 1970s.[119][120] The National Weather Service however, has rejected naming winter storms.[121] Despite this, the Free University of Berlin kept the name "Nemo", referring to the storm by that name on their weather maps.[40] Other names for the storm include the "Blizzard of 2013" (or the "Blizzard of '13") and "Blizzard 2013".[7][122]

TRB's National Cooperative Highway Research Program (NCHRP) Research Report 840: A Watershed Approach to Mitigating Stormwater Impacts provides a practical decision-making framework that will enable state departments of transportation (DOTs) to identify and implement offsite cost-effective and environmentally beneficial water quality solutions for stormwater impacts when onsite treatment and/or mitigation is not possible within the right-of-way.

RGENs are derived from the type II CRISPR (clusters of regularly interspaced palindromic repeats)/Cas (CRISPR-associated) system, an adaptive immune response in bacteria and archaea (Wiedenheft et al. 2012). Cas9, the protein component derived from Streptococcus pyogenes, forms an active nuclease when complexed with transactivating CRISPR RNA (tracrRNA) and CRISPR RNA (crRNA) (Jinek et al. 2012), which are transcribed from the CRISPR sequence encoded in the bacterial genome. This ribonucleoprotein protects host cells from invading phages or plasmids by recognizing and cleaving the DNA sequence corresponding to the crRNA sequence. Recently, we and others have exploited this system to induce site-specific DSBs, thereby modifying genomes in a targeted manner in cells and organisms (Chang et al. 2013; Cho et al. 2013a,b; Cong et al. 2013; Ding et al. 2013; Gratz et al. 2013; Hwang et al. 2013; Jiang et al. 2013; Jinek et al. 2013; Mali et al. 2013b; Shen et al. 2013; Wang et al. 2013).

Comparison of guide RNA structure. Mutation frequencies of the RGENs reported in Fu et al. (2013) were measured at on-target and off-target sites using the T7E1 assay. K562 cells were cotransfected with the Cas9-encoding plasmid and the plasmid encoding GX19 sgRNA or GGX20 sgRNA. Off-target sites (OT1-3, etc.) are labeled as in Fu et al. (2013).

In this study, we showed that RGENs do indeed induce off-target mutations at sites with a single-base mismatch. We found, however, that RGENs efficiently discriminate on-target sites from off-target sites that differ by only two bases. Furthermore, exome sequencing showed that no off-target mutations were present in four clonal populations of mutant cells. Our results suggest that one could avoid or minimize off-target effects of RGENs by choosing unique target sites that do not have any homologous sequences elsewhere in the genome, a strategy we had used to avoid off-target effects of TALENs (Kim et al. 2013a). We also found that the structure and composition of guide RNA can be modified to reduce off-target mutations. We cannot rule out the possibility that the 11 RGENs we created in this study induce off-target mutations at sites not examined here, which could be revealed by deep sequencing at other less-homologous candidate sites or by whole genome sequencing. In addition, in vitro selection of cleavage sites (Pattanayak et al. 2011) or the IDLV capture approach (Gabriel et al. 2011) used for the identification of ZFN off-target sites may reveal cryptic sites cleaved by RGENs, although it is difficult to imagine that RGENs recognize off-target sites that are not complementary to crRNA sequences at an appreciable level. It is also worth noting that RGENs cleave DNA much less discriminatingly in vitro than they do in cells (Fig. 1C), limiting the in vitro selection method. 076b4e4f54


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