A faster way to make GFP-based biosensors: Two new transposons for creating multicolored libraries of fluorescent fusion proteins

BACKGROUND: There are now several ways to generate fluorescent fusion proteins by randomly inserting DNA encoding the Green Fluorescent Protein (GFP) into another protein's coding sequence. These approaches can be used to map regions in a protein that are permissive for GFP insertion or to create novel biosensors. While remarkably useful, the current insertional strategies have two major limitations: (1) they only produce one kind, or color, of fluorescent fusion protein and (2) one half of all GFP insertions within the target coding sequence are in the wrong orientation. RESULTS: We have overcome these limitations by incorporating two different fluorescent proteins coding sequences in a single transposon, either in tandem or antiparallel. Our initial tests targeted two mammalian integral membrane proteins: the voltage sensitive motor, Prestin, and an ER ligand gated Ca(2+ )channel (IP(3)R). CONCLUSIONS: These new designs increase the efficiency of random fusion protein generation in one of two ways: (1) by creating two different fusion proteins from each insertion or (2) by being independent of orientation

Ames collaborative study of cosmic-ray neutrons. 2: Low- and mid-latitude flights

Progress of the study of cosmic ray neutrons is described. Data obtained aboard flights from Hawaii at altitudes of 41,000 and 45,000 feet, and in the range of geomagnetic latitude 17 N less than or equal to lambda less than or equal to 21 N are reported. Preliminary estimates of neutron spectra are made

Age, Growth, Relative Abundance, and Scuba Capture of a New or Recovering Spawning Population of Lake Sturgeon in the Lower Niagara River, New York

The objective of our study was to collect age, growth, and catch-per-unit-effort information from a new or recovering population of lake sturgeon Acipenser fulvescens in the lower Niagara River, New York. From July 1998 through August 2000, we captured 67 lake sturgeon by use of gill nets, baited setlines, and scuba diving. Active capture by scuba divers (1.50 fish/ h) was much more effective than passive capture with gill nets (0.07 fish/h) and setlines (0.06 fish/h). Eggs of Chinook salmon Oncorhynchus tshawytscha were more effective as setline bait than were alewives Alosa pseudoharengus, but neither bait differed in effectiveness from rainbow smelt Osmerus mordax. Ages of captured lake sturgeon ranged from 1 to 23 years; 47 of the 61 aged fish were younger than age 10. Strong relationships were found between weight, W, and length, L (W = 0.0000005 L3.5564: R2 = 0.977) and between L and age (L = 394.05.log e[age] + 248.77; R2 = 0.878). The lake sturgeon population in the lower Niagara River is probably small relative to its historic abundance. This naturally reproducing population should remain listed as threatened by New York State, and commercial and recreational fisheries should remain closed so that the population can rebuild adult numbers and reproductive potential

Late Pleistocene Interactions of East and West Antarctic Ice-Flow Regimes: Evidence from the McMurdo Ice Shelf

We present new interpretations of deglaciation in McMurdo Sound and the western Ross Sea, with observationally based reconstructions of interactions between East and West Antarctic ice at the last glacial maximum (LGM), 16 000, 12 000, 8000 and 4000 sp. At the LGM? East Antarctic ice from Mulock Glacier split, one branch turned westward south of Ross Island but the other branch rounded Ross Island before flowing southwest into McMurdo Sound. This flow regime, constrained by an ice saddle north of Ross Island, is consistent with the reconstruction of Stuiver and others (1981a). After the LGM, grounding-line retreat was most rapid in areas with greatest water depth, especially along the Victoria Land coast. By 12 000 sp, the ice-now regime in McMurdo Sound changed to through-flowing Mulock Glacier ice, with lesser contributions from Koettlitz, Blue and Ferrar Glaciers, because the former ice saddle north of Ross Island was replaced by a dome. The modern flew regime was established similar to 4000 BP. Ice derived from high elevations on the Polar Plateau but now stranded on the McMurdo Ice Shelf, and the pattern of the Transantarctic Mountains erratics support our reconstructions of Mulock Glacier ice rounding Minna Bluff but with all ice from Skelton Glacier ablating south of the bluff. They are inconsistent with Drewry\u27s (1979) LGM reconstruction that includes Skelton Glacier ice in the McMurdo-Sound through-flow. Drewry\u27s (1979) model closely approximates our results for 12 000-4000 BP. Ice-sheet modeling holds promise for determining whether deglaciation proceeded by grounding-line retreat of an ice sheet that was largely stagnant, because it never approached equilibrium flowline profiles after the Ross Ice Shelf, grounded, or of a dynamic ice sheet with flowline profiles kept low by active ice streams that extended northward from present-day outlet glaciers after the Ross Ice Shelf grounded

Adaptation of the DEMQOL-Proxy for routine use in care homes: a cross sectional study of the reliability and validity of DEMQOL-CH

Objective: To investigate the routine use of a measure of quality of life (QoL) in care homes and assess its psychometric properties when used by care staff Design: A cross-sectional two-phase study. Setting and participants: Data were collected from care staff in seven care homes in East Sussex, England. Method: Phase 1: The ability of care staff from two care homes to use the DEMQOL-Proxy without interviewer-administration was assessed using agreement analysis between a selfand interviewer-administered version of the instrument. Based on these findings, DEMQOLProxy was adapted into a new version, DEMQOL-CH, for use as a self-administered instrument in care homes. We assessed agreement between the new DEMQOL-CH and DEMQOL-Proxy to ensure DEMQOL-CH was used correctly. Phase 2: A preliminary assessment of the psychometric properties of DEMQOL-CH when used routinely was completed in a further five care homes. Results: Phase 1: Nineteen care staff from two care homes completed QoL measurements for residents. Systematic error was identified when staff self-completed the DEMQOL-Proxy without an interviewer. We modified the DEMQOL-Proxy to create DEMQOL-CH; this reduced the error, producing a version that could be used more accurately by care staff. Phase 2: Eleven care staff from five care homes rated resident QoL routinely. DEMQOL-CH showed acceptable psychometric properties with satisfactory reliability and validity and a clear factor structure. Conclusions: The research presents positive preliminary data on the acceptability, feasibility and performance of routine QoL measurement in care homes using an adapted version of DEMQOL-Proxy, the DEMQOL-CH. Results provide evidence to support the concept that routine measurement of QoL may be possible in care homes. Research is needed to refine and test the methodology and instrument further, and to explore the potential for benefits to residents, staff, and care homes in larger and more representative populations

Psychometric properties and feasibility of use of dementia specific quality of life instruments for use in care settings: a systematic review

Background: Over 400,000 people live in care home settings in the UK. One way of understanding and improving the quality of care provided is by measuring and understanding the quality of life (QoL) of those living in care homes. This review aimed to identify and examine the psychometric properties including feasibility of use of dementia-specific QoL measures developed or validated for use in care settings. Design: Systematic review. Methods: Instruments were identified using four electronic databases (PubMed, PsycINFO, Web of Science, and CINAHL) and lateral search techniques. Searches were conducted in January 2017. Studies which reported on the development and/or validation of dementia specific QoL instruments for use in care settings written in English were eligible for inclusion. The methodological quality of the studies was assessed using the COSMIN checklist. Feasibility was assessed using a checklist developed specifically for the review. Results: Six hundred and sixteen articles were identified in the initial search. After de-duplication, screening and further lateral searches were performed, 25 studies reporting on 9 dementia-specific QoL instruments for use in care home settings were included in the review. Limited evidence was available on the psychometric properties of many instruments identified. Higher-quality instruments were not easily accessible or had low feasibility of use. Conclusions: Few high-quality instruments of QoL validated for use in care home settings are readily or freely available. This review highlights the need to develop a well-validated measure of QoL for use within care homes that is also feasible and accessible

Using emergency department syndromic surveillance to investigate the impact of a national vaccination program: A retrospective observational study

BackgroundRotavirus infection is a common cause of gastroenteritis in children worldwide, with a high mortality burden in developing countries, particularly during the first two years of life. Rotavirus vaccination was introduced into the United Kingdom childhood vaccination schedule in July 2013, with high coverage (>90%) achieved by June 2016. We used an emergency department (ED) syndromic surveillance system to assess the impact of the rotavirus vaccination programme, specifically through the demonstration of any immediate and continuing impact on ED gastroenteritis visits in England.MethodsThis retrospective, observational study used syndromic surveillance data collected from 3 EDs in the two years before (July 2011-June 2013) and 3 years post (July 2013-June 2016) introduction of rotavirus vaccination. The weekly levels of ED visits for gastroenteritis (by age group and in total) during the period before rotavirus vaccination was first described alongside the findings of laboratory surveillance of rotavirus during the same period. An interrupted time-series analysis was then performed to demonstrate the impact of rotavirus vaccination introduction on gastroenteritis ED visit levels.ResultsDuring the two years before vaccine introduction ED visits for gastroenteritis in total and for the 0-4 years age group were seen to rise and fall in line with the seasonal rotavirus increases reported by laboratory surveillance. ED gastroenteritis visits by young children were lower in the three years following introduction of rotavirus vaccination (reduced from 8% of visits to 6% of visits). These attendance levels in young children (0-4years) remained higher than in older age groups, however the previously large seasonal increases in children were greatly reduced, from peaks of 16% to 3-10% of ED visits per week.ConclusionsED syndromic surveillance demonstrated a reduction in gastroenteritis visits following rotavirus vaccine introduction. This work establishes ED syndromic surveillance as a platform for rapid impact assessment of future vaccine programmes