- Last 7 days
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writingball.blogspot.com writingball.blogspot.com
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KMC vs. KMM by [[Richard Polt]]
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theconversation.com theconversation.com
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Die von Waldbränden außerhalb der Tropen verursachten Emissionen haben sich seit 2001 fast verdreifacht. Weltweit haben die Emissionen durch Waldbrände in dieser Zeit um 60% zugenommen. Ursache dafür ist die Kombination von heißerem und trockenerem Wetter mit dem schnelleren Wachstum der Wälder durch die höheren Temperaturen. Die Wälder können durch die Brände jahrzehntelang zu Emittenten werden. Damit ist die Funktion der Wälder als Kohlenstoffsenken gefährdet. Das bedeutet auch, dass sie andere anthropogene Emissionen weniger kompensieren und die Fähigkeit verlieren, nach einem Überschreiten der 1,5°-Grenze C0<sub>2</sub> aus der Atmosphäre zu entfernen. Außerdem müssten diese von Menschen verursachten Emissonen den C0<sub>2</sub>-Budgets der Nationalstaaten zugeordnet werden.
Tags
- author:: Crystal A. Kolden
- date::2024-10-17
- increasing risk of wildfires
- Boreale Wälder
- author:: Matthew W. Jones
- author: Stefan H. Doerr
- date:: 2024-10-17
- Schwächung der terrestrischen Kohlenstoffsenken
- Global rise in forest fire emissions linked to climate change in the extratropics
- 2001-2023
Annotators
URL
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- Sep 2024
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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GMR-Gal4(II)
DOI: 10.1128/MCB.00195-17
Resource: Bloomington Drosophila Stock Center (RRID:SCR_006457)
Curator: @maulamb
SciCrunch record: RRID:SCR_006457
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- Aug 2024
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www.gov.ca.gov www.gov.ca.gov
- Jul 2024
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en.wikipedia.org en.wikipedia.org
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https://en.wikipedia.org/wiki/Matthew_effect
The Matthew effect of accumulated advantage, sometimes called the Matthew principle, is the tendency of individuals to accrue social or economic success in proportion to their initial level of popularity, friends, and wealth. It is sometimes summarized by the adage or platitude "the rich get richer and the poor get poorer". The term was coined by sociologists Robert K. Merton and Harriet Zuckerman in 1968 and takes its name from the Parable of the Talents in the biblical Gospel of Matthew.
related somehow to the [[Lindy effect]]?
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docs.google.com docs.google.com
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Please sign these letters to legislators, telling them that misguided AI laws will hurt startups and small companies and discourage AI innovation and investment in California.AI offers tremendous benefits, but many fear AI and worry about potential harm and misuse. These are valid concerns for everyone, including legislators, but laws that promote safe and equitable AI should be fact-based, straightforward, and universally applied. Legislators in Sacramento are considering two proposals, AB 2930 and SB 1047, that would impose costly and unpredictable burdens on AI developers, including anticipating and preventing future harmful uses of AI. Though well-intended, these bills will dampen and inhibit innovation, permanently embed today’s AI leaders as innovation gatekeepers, and drive investment and talent to other states and countries.
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en.wikipedia.org en.wikipedia.org
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The g0v movement, or g0v, (pronounced gov-zero /ɡʌvziroʊ/) is an open source, open government collaboration started by Chia-liang Kao ("clkao"), ipa, kirby and others in late 2012 in Taiwan.
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github.com github.com
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The Computational Democracy Project
We bring data science to deliberative democracy, so that governance may better reflect the multidimensionality of the public's will.
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Input Crowd, Output Meaning
Polis<br /> https://pol.is/home
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www.youtube.com www.youtube.com
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www.reddit.com www.reddit.com
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https://www.reddit.com/r/movies/comments/kzc5o3/are_gene_hackman_characters_in_the_conversation/<br /> Are Gene Hackman characters in The Conversation (1974) and Enemy of the State (1998 ) connected?
I want to circle back to this as I've had this idea since I saw Enemy of the State as a rough cut.
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moshi.chat moshi.chat
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https://moshi.chat/
Moshi is an experimental conversational AI.
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- May 2024
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www.theguardian.com www.theguardian.com
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Das globale Durchschnittseinkommen wird bei der jetzt zu erwartenden globalen Erhitzung 2050 fast um ein Fünttel niedriger sein als ohne Erhitzung. Die (nicht mehr zu vermeidenden) Einbußen durch die Erhitzung bis 2050 sind sechsmal so hoch wie die einer Begrenzung des Temperaturanstiegs auf 2°. 2050 ist einer neuen Studie zufolge mit Klimaschäden von etwa 38 Bllionen Dollar zu rechnen. Bis 2100 wird es in einem Business-as-usual-Szenario zu Einkommensverlusten von mehr als 60% kommen. https://www.theguardian.com/environment/2024/apr/17/climate-crisis-average-world-incomes-to-drop-by-nearly-a-fifth-by-2050
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www.theguardian.com www.theguardian.com
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2021 haben sich viele, vor allem britische, Banken zu sogenannten Glasgow Financial Alliance for Net Zero (GFANZ) zusammengeschlossen. Ein Bericht der Organisation Reclaim Finance weist nach, dass über 50 von ihnen weiterhin massiv Investitionen in fossile Energien finanziert haben.
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www.theguardian.com www.theguardian.com
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Der Artikel im Guardian stellt eine neue Studie dar, aus der hervorgeht, wie viel von der bereits existierenden Infrastruktur zur Förderung fossiler Brennstoff stillgelegt werden muss, um das 1,5° Ziel zu erreichen. Dabei geht die Autoren davon aus, dass man CO2 nicht realistisch wieder aus der Atmosphäre entfernen kann, und dass das 1,5° Ziel also nur zu erreichen ist, wenn nicht zu viel emittiert wird. Diese Studie fordert das Gegenteil der Planungen der fossilen Industrien, über der über die der Guardian gerade berichtet hatte. Der Artikel ist auch bemerkenswert, weil er auf eine Reihe weiterer wichtiger Studien zu fossilen Lagerstätten verweist.
Tags
- 2022-05-17
- expert: Kelly Trout
- institution: Beyond Oil and Gas Alliance
- topic: carbon removal
- treaty: energy charter treaty
- fossil fuels
- expert: Maeve O'Connor
- variable: emissions of developed reserves
- institution: Carbon Tracker
- Climate emergency
- fossil expansion
- expert: Daniel Welsby
- variable: emissions of known reserves
- expert: Greg Muttitt
Annotators
URL
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- Apr 2024
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www.theguardian.com www.theguardian.com
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Im Februar setzen sich die Messungen von Rekordtemperaturen fort. Dabei stellt seit über einem halben Jahri die Zahl dieser Meldungen selbst bisherige Rekorde ein. In den ersten beiden Wochen wurden an 140 Stellen der Erde die höchsten Monats-temperaturen der Aufzeichnunggsgeschicht festgestellt. Noch beunruhigender sind die Anomaliee bei den Meerestemperaturen. Guardian-Artikel mit SInfografiken und tatements von Klimawissenschaftler:innen, die von dieser Entwicklung alarmiert sind. https://www.theguardian.com/environment/2024/feb/17/february-on-course-to-break-unprecedented-number-of-heat-records
Tags
- The Nature Conservancy
- by: Jonathan Watts
- global
- Zeke Hausfather
- Joel Hirschi
- El Niño
- 2024-02-17
- UK National Oceanography Centre
- Michael Lowry
- topic: temperature records
- Berkeley Earth
- Katharine Hayhoe
- China
- sea surface temperature
- Francesca Guglielmo
- Copernicus
- Maximiliano Herrera
- Morocco
- global heating
Annotators
URL
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www.zoni.edu www.zoni.edu
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Networking 1B (0.5)
It's Intro to Networking 1B (0.5)
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www.derstandard.at www.derstandard.at
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www.theguardian.com www.theguardian.com
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In Norditalien setzt sich die Dürre-Situation des vergangenen Jahres fort, weil - wie in Frankreich - im Winter lange kein Regen gefallen ist. Der Wasserstand des Po ist extrem niedrig. Maßnahmen zur Kontrolle des Wasserverbrauchs sind unter der Rechtsregierung noch schwieriger durchzusetzen als in der Regierungszeit Draghis.
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- Mar 2024
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www.derstandard.de www.derstandard.de
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Das österreichische Klimaschutzministerium stellt in diesem Jahr 150 Millionen Euro Subventionen für den Ausbau erneuerbarer Energien zur Verfügung. Dabei geht es im Wesentlichen um gewerblichen Nutzung. https://www.derstandard.de/story/3000000212023/klimaministerium-stellt-heuer-150-mio-euro-f252r-214kostromausbau-bereit
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www.derstandard.at www.derstandard.at
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Dass Erneuerbare-Wärme-Gesetz wird gegenüber dem ursprünglichen Gewessler-Entwurf deutlich verwässert: Auf das Stillegungsgebaut für bestehende Öl- und Gasheizungen wird ebenso verzichtet wie auf das Zentralisierungsgebot. Es werden dafür viele Förderungen angeboten. https://www.derstandard.at/story/3000000191501/regierung-streicht-komplettausstieg-bei-oel-und-gas
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- Feb 2024
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www.liberation.fr www.liberation.fr
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Viele – möglicherweise alle – Populationen von Eisbären drohen bis zum Ende des Jahrhunderts zu verschwinden, wenn das Verfeuern fossiler Brennstoffe nicht gestoppt wird. Interview mit dem Biologen Geoff York zur Bewohnbarkeit der Arktis für Eisbären, aber auch für andere Arten und den Menschen. https://www.liberation.fr/environnement/biodiversite/pour-sauver-les-ours-polaires-la-seule-solution-est-darreter-de-bruler-des-energies-fossiles-20240217_LZ2BEPEJXJHRHKWINWHE6Q57XA/
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Annotators
URL
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www.derstandard.de www.derstandard.de
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images.lambethpalacelibrary.org.uk images.lambethpalacelibrary.org.uk
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images.lambethpalacelibrary.org.uk images.lambethpalacelibrary.org.uk
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Robert Grosseteste, Bishop of Lincoln (face rubbed), in mitre and red cope, with crosier, seated on left speaks to a seated group of five people, mostly women. Tree on right; large bird with long beak at top.
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medieval.bodleian.ox.ac.uk medieval.bodleian.ox.ac.uk
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https://medieval.bodleian.ox.ac.uk/catalog/work_4353
Manuscript copies of Chasteau d'amour (The Castle of Love) by Robert Grosseteste (1175? - 1253) in the Oxford Libraries.
No copies available digitally as of 2024-02-16
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- Jan 2024
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www.repubblica.it www.repubblica.it
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Zwei der Reports, die zum Weltwirtschaftsforum 2024 publiziert wurden, betonen die Bedeutung von Risiken, die mit der globalen Erhitzung, der Zerstörung der Biodiversität und der lebenserhaltenden Systeme des Planeten verbunden sind. Der Artikel der Repubblica zählt klimapolitisch wichtige Ereignisse des Jahres 2024 auf.https://www.repubblica.it/green-and-blue/2024/01/17/news/world_economic_forum_2024_cambiamento_climatico-421899576/
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www.repubblica.it www.repubblica.it
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Das EU-Parlament hat die Anti-Greenwashing-Direktive abschließend gebilligt. Unter anderem dürfen Produkte nicht mehr als nachhaltig gelabelt werden, die durch CO<sub>2</sub>-Zertifikate für Kompensationen angeblich klimaneutral gemacht werden. https://www.repubblica.it/green-and-blue/2024/01/17/news/ue_greenwashing_messaggi_ingannevoli_approvata_legge-421900621/
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www.derstandard.at www.derstandard.at
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Der Umsatz des österreichischen Ölfeldausrüsters SBO stieg 2023 um 17%. SBO profilierte vor allem vom Wachstum der Ölmärkte. https://www.derstandard.at/story/3000000203464/sbo-2023-mit-rekordumsatz-von-585-millionen-euro
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www.derstandard.at www.derstandard.at
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45% der CO<sub>2</sub>-Emissionen entstehen durch die Produktion von Materialien. Der Standard berichtet über eine Podiumsdiskussion in Wien, aus der hervorgehen, das Recycling allein die ökologischen Probleme, die dadurch entstehen, nicht löst. https://www.derstandard.at/story/3000000203446/warum-recycling-kein-ausweg-aus-der-materialschlacht-ist
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www.theguardian.com www.theguardian.com
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Der grönländische Eisschild verliert aufgrund der globalen Erhitzung 30 Millionen Tonnen Eis pro Stunde und damit 20% mehr als bisher angenommen. Manche Forschende fürchten, dass damit das Risiko eines Kollaps des Amoc größer ist als bisher angenommen. Der Eisverlust ist außerdem relevant für die Berechnung des Energie-Ungleichgewichts der Erde durch Treibhausgas-Emissionen. https://www.theguardian.com/environment/2024/jan/17/greenland-losing-30m-tonnes-of-ice-an-hour-study-reveals
Tags
- AMOC
- Antarctic calving loss rivals ice-shelf thinning
- Jet Propulsion Lab
- Andrew Shepherd
- cryosphere
- Tim Lenton
- Chad Greene
- Ubiquitous acceleration in Greenland Ice Sheet calving from 1985 to 2022
- Greenland
- 2024-01-17
- Mass balance of the Greenland and Antarctic ice sheets from 1992 to 2020
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URL
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www.liberation.fr www.liberation.fr
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20% der Schneemasse auf der Nordhalbkugel bedecken Gebiete, die im Winter meist wärmer sind als 8°. In diesen Gebieten hat die Schneedecke in den letzten Jahrzehnten bereits deutlich abgenommen. Für ihre Zukunft ist jedes Zehntelgrad mehr oder weniger Erhitzung entscheidend. Der Verlust der Schneedecke führt zu Problemen bei der Wasserversorgung etwa der Donau und des Mississippi. https://www.liberation.fr/environnement/y-aura-t-il-encore-de-la-neige-en-2050-20240117_UPOQVWROIZEBVDQRD5JBRA4EH4/
Mehr zur selben Studie: https://hypothes.is/search?q=tag%3A%22Evidence%20of%20human%20influence%20on%20Northern%20Hemisphere%20snow%20loss%22
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www.repubblica.it www.repubblica.it
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Die Desinformation zur globalen Erhitzung hat sich von der Klimaleugnung hin zum Säen von Zweifeln an möglichen Lösungen verschoben. Einer neuer Studie zufolge sind wichtige Strategien auf Youdas Tube das Herunterspielen der negativen Konsequenzen, Erzeugen von Misstrauen in die Klimaforschung und vor allem die Behauptung, dass vorhandene technische Lösungen nicht praktikabel sind. Außerdem werden Verschwörungstheorien wie die vom Grand Reset bemüht. https://www.repubblica.it/green-and-blue/2024/01/17/news/negazionismo_climatico_youtube-421894897/
Studie: https://counterhate.com/wp-content/uploads/2024/01/CCDH-The-New-Climate-Denial_FINAL.pdf
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Die 20 größten Unternehmen der Welt sind in den vergangenen Jahren noch mächtiger geworden. Das zeigt eine neue Studie mehrerer NGOs. Diese Unternehmen können Marktpreise und Produktlinien diktieren, weil sie fast keine Konkurrenz haben. Sie betreiben intensives Lobbying. https://taz.de/Warnung-vor-Macht-der-Grossunternehmen/!5986399/
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Der deutsche Finanzminister Lindner plant Kürzungen bei ursprünglich für die Verkehrswende, vor allem für die Verbesserung der Schieneninfrastruktur vorgesehenen Geldern. NGOs und Verbände protestieren dagegen. Beim Straßenbau sind kaum Kürzungen geplant. https://taz.de/Sparplaene-fuer-den-Bundeshaushalt/!5986400/
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www.liberation.fr www.liberation.fr
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www.liberation.fr www.liberation.fr
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Wieder was auch beschäftigt sich damit, dass isch die Wahrnehmung des Wetters verändert, weil man sich an die geänderten Bedingungen anpasst und die früheren Verhältnisse vergisst Punkt Menschen ohne 30 haben in Frankreich in der regelkrankheiten Winter mehr erlebt und halten ihr fehlen deshalb auch für selbstverständlich. https://www.liberation.fr/environnement/climat/avec-le-rechauffement-des-temperatures-les-hivers-perdus-entre-amnesie-et-nostalgie-20240117_RINBQOXOLRDU7GZ27UMHFDBH3U/
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Der Ausbau der Windenergie in Deutschland hat sich unter der Ampelkoalition beschleunigt. Es werden aber im Augenblick nur etwa halt so viele Anlagen gebaut, wie es nach den Plänen der Bundesregierung nötig wäre. In den südlichen Bundesländern ist der Ausbau viel zu langsam. Genehmigungsverfahren z.B. für den Transport dauern zu lange. https://taz.de/Ausbau-der-Windenergie/!5983011/
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www.handelsblatt.com www.handelsblatt.com
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Die europäische Solar-Industrie ist in einer schweren Krise, weil chinesische Unternehmen mit wesentlich billigeren Modulen auf den europäischen Markt drängen. Die Subventionen der USA für die eigene Industrie verstärkt diesen Druck. Wenn die deutsche Regierung ihre Subventionspolitk nicht ändert, wird der Hersteller Meyer Burger sein deutsches waren schließen und nur noch in den USA produzieren. https://www.handelsblatt.com/unternehmen/energie/erneuerbare-energie-solarausruester-meyer-burger-droht-mit-schliessung-der-deutschen-modulproduktion/100007780.html
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www.theguardian.com www.theguardian.com
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Eine neue Studie kommt zu dem Ergebnis, dass die Haltung zu fünf großen Krisen das Wahlverhalten der Europäer:innen in diesem Jahr bestimmen wird: der Klimakrise, der Migrationskrise, der Wirtschaftskrise und Inflation, dem Ukraine-Krieg und Covid. Klimakrise und Migration hätten, wie schon bei den Wahlen in der Niederlanden, ide größte Kraft Wähler zu mobilisieren. Die Autor:innen sprechen von einem "Clash zweier 'Extinction rebellions'". Als wichtigste Krisen werden im Durchschnitt der europäischen Länder die Klimakrise und dann Covid bewertet.
Report: https://ecfr.eu/publication/a-crisis-of-ones-own-the-politics-of-trauma-in-europes-election-year/
Tags
- Mark Leonard
- A Crisis of One’s Own: the Politics of Trauma in Europe’s Election Year
- 2024 European elections
- Centre for Liberal Strategies
- European Council on Foreign Relations (ECFR)
- Ivan Krastev
- 2024-01-17
Annotators
URL
theguardian.com/world/2024/jan/17/crises-have-split-european-voters-into-five-tribes-survey-suggests -
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www.liberation.fr www.liberation.fr
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27 Aktionäre von Shell unterstützen eine Resolution, die den Konzern auffordert, das Pariser Abkommen einzuhalten. Sie repräsentieren 3,4% des Kapitals der Firma. Dieselbe Resolution hatte bei TotalEnergies die Unterstützung von 30% des Kapitals gefunden. Im vergangenen Jahr hatte Shell seine ohnehin minimalen Dekarbonisierungsziele revidiert und angekündigt, die Ölproduktion bis 2030 stabil zu halten. https://www.liberation.fr/environnement/pollution/industries-fossiles-chez-shell-des-actionnaires-activistes-font-pression-pour-de-meilleurs-objectifs-climatiques-20240116_HKBWVGK5CVFMDACJBNKNMP4E2A/
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Annotators
URL
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www.repubblica.it www.repubblica.it
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- Dec 2023
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www.liberation.fr www.liberation.fr
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In Frankreich hat der flussschau zum zweiten Mal innerhalb weniger Monate Überschwemmungen verursacht, auch vor zwei Jahren gab es dort Überflutungen. AnwohnerInnen beobachten eine deutliche Zunahme von Überschwemmungen. https://www.liberation.fr/societe/la-charente-deborde-letat-de-catastrophe-naturelle-va-etre-demande-20231217_WIWWRVFTDJHXPA4OBEFXT3FWOM/
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www.legifrance.gouv.fr www.legifrance.gouv.fr
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Le comité départemental de suivi de l'école inclusive encourage le développement des actions de formation croisée en matière d'école inclusive et de coopération. Il en dresse le bilan
Tags
Annotators
URL
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www.theguardian.com www.theguardian.com
Tags
- NGO: Energy and Climate Intelligence Unit
- NGO: Leave It in the Ground Initiative
- 2023-08-17
- expert: David Tong
- actor: Adnoc
- expert: Kjell Kühne
- Adnoc
- actor: Sultan Al Jaber
- expert: Gareth Redmond-King
- process: methane reduction
- country: UAE
- topic: Methane emissions
- event: COP28
- NGO: Oil Change International
Annotators
URL
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- Nov 2023
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www.theguardian.com www.theguardian.com
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Auf den Öl- und Gasfeldern der Vereinigten Arabischen Emirate, darunter vielen, die der staatlichen Gesellschaft Adnoc gehören, wurde in den vergangenen 20 Jahren in großem Umfang routinemäßig Gas abgefackelt, was zu hohen Methanemissionen führt. Die Emirate hatten sich verpflichtet, das Abfackeln schnell zu reduzieren. Die dieser Selbstverpflichtung krass widersprechende Praxis gilt bei NGO als weiterer Beleg dafür, dass Selbstverpflichtungen der Fossilindustrie nicht getraut werden kann. https://www.theguardian.com/environment/2023/nov/17/cop28-host-uae-breaking-its-own-ban-on-routine-gas-flaring-data-showsactor
Tags
- 2023-11-17
- topic: gas flaring
- by: Damian Carrington
- expert: Hubert Thieriot
- expert: Zubin Bamji
- actor: Sultan Al Jaber
- actor: Dubai Petroleum
- NGO: Kick Big Polluters Out
- expert: Pascoe Sabido
- topic: Methane emissions
- institution: Centre for Research on Energy and Clean Air (Crea).
- actor: Adnoc
- expert: Paul Balcombe
- NGO: Corporate Europe Observatory
- country: UAE
- topic: LNG
- institution: World Bank
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URL
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Die Klimaökonomin Claudia Kemfert hat wegen der Entscheidung des Bundesverfassungsgerichts, die Verwendung von Corona -Rücklagen für den Klima- und Transformationsfonds zu untersagen, von einem schwarzen Tag für den Klimaschutz gesprochen. Sie schlägt vor, den Klimanotstand auszurufen oder fossile Subventionen massiv zu kürzen. https://taz.de/Nach-Karlsruher-Urteil-zum-Bundesetat/!5969938/
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www.theguardian.com www.theguardian.com
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paultough.com paultough.com
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key factor behind academic perseverance was students’ academic mindset — the attitudes and self-perceptions
Academic Mindset = attitudes + self-perceptsion
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act gritty
Interesting distinction - being gritty vs. acting gritty
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all students are more likely to demonstrate perseverance if the school or classroom context helps them develop positive mindsets and effective learning strategies.”
How can we. help students develop positive mindsets and effective learning strategies?
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If they hear the message that a failure is a final verdict on their ability, they may well give up and pull back from school. But if instead they get the message that a failure is a temporary stumble, or even a valuable opportunity to learn and improve, then that setback is more likely to propel them to invest more of themselves in their education.
Are these the only two types of messages a student can receive in response to their failure?
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She was particularly interested in what she called the “narrative” that exists within each school with regard to success and failure — the messages, subtle and not so subtle, that students receive when they fai
What is the "narrative" at your SL school that a student receives when they fail?
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17. Messages
An examination of the messages teachers send to students - intentionally or not.
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institutional structures affect individual behavior and, specifically, how certain educational structures — like school funding mechanisms, teacher contracts, or patterns of segregation — might incline students toward success or failure.
So Farrington was looking at the issue from a individual level (psych), as well as more societal/institutional level (sociology)
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- Oct 2023
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www.theguardian.com www.theguardian.com
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www.theguardian.com www.theguardian.com
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www.derstandard.at www.derstandard.at
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www.liberation.fr www.liberation.fr
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- Sep 2023
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support.apple.com support.apple.com
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Find Alarm now replaces Get All Alarms, retrieving all alarms or only those which match filter criteria
Can't believe what I long called "by far and away Siri's most important function in my life" must now be intentionally configured and (non-verbally) triggered in the form of a Siri Shortcut.
Thank you.
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Transcribe Audio generates text from an audio file
This technically works, yes, but - as I noticed on the very first beta build - the actual output has an absurdly low character(?) limit, after which it simply cuts off.
Tags
Annotators
URL
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www.apple.com www.apple.com
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I think it is absolutely absurd that Apple still(?) chooses to publish "complete" ""feature lists"" in Portable Document Format, of all motherfucking things, but - as always - thank fucking God for mr Hypo Thesis over here!
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www.derstandard.de www.derstandard.de
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support.apple.com support.apple.com
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gently nudging the table your iPhone is on
EXCUSE ME????
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support.apple.com support.apple.com
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Passengers must have an iPhone with iOS 17 or later, but don’t need to have an Apple Music subscription.
Well, now I've confirmed the most crucial answer I needed answering about iOS 17...
...now just to figure out how I'm going to make this happen.
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www.theguardian.com www.theguardian.com
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Die Ölpreise steigern deutlich an, vor allem aufgrund von steigender Nachfrage in China (dem größten Öl-Importeur) und Drosselung der Produktion in Saudi-Arabien und Russland. Hintergrund dieser Verknappung ist auch die Prognose der IEA, die Nachfrage nach Öl werde vor 2030 erstmals sinken. https://www.theguardian.com/business/2023/sep/17/global-inflation-fears-as-oil-price-rises-towards-100-a-barrel
-
-
www.theguardian.com www.theguardian.com
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I'm Vorfeld des UN Climate Ambition Summit am Mittwoch nahmen zwischen 50.000 und 75.000 Menschen (nach Angaben der Organisierenden) in New York am "March to end fossil fuels" teil. Damit war er die größte Klimakundgebung in den USA seit dem Beginn der Pandemie. Alexandria Ocasio-Cortez forderte in einer Rede das Ende aller fossilen Projekte in den USA und eine demokratische Emergie-Wirtschaft. https://www.theguardian.com/environment/2023/sep/17/march-to-end-fossil-fuels-new-york-city
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- Jun 2023
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gaily
happily or brightly 歡樂地，喜氣洋洋地；閃亮地，明亮地
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mottled
covered with areas of different colours that do not form a regular pattern 雜色的；斑駁的
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elapsed
If time elapses, it goes past.（時間）流逝，過去
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pierced
to go into or through something, making a hole in it using a sharp point 刺穿，刺透，刺破
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slumbe
sleep
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- Mar 2023
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Great sorrow filled my heart on hearing this,because I knew of people of great worth,who in that Borderland suspended were.
He still not understand that God is justice itself.
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against me this one seemed to be advancingwith head erect and with such raging hunger,that even the air seemed terrified thereby—and of a she-Wolf, which with every lustseemed in her leanness laden, and had caused 50many ere now to lead unhappy lives.
This imply that lust is the most difficult one for humans to overcome.
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Art thou that Virgil, then, that fountain-headwhich poureth forth so broad a stream of speech?”
Why does Dante choose Virgil?
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When half way through the journey of our lifeI found that I was in a gloomy wood,because the path which led aright was lost.
This is also Dante reminding himself.
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I had his hair wrapped round my hand already,and more than one shock had I plucked from him,while he was barking, with his eyes turned down
Now Dante fully accepted the fact that God is justice. Those people deserve to be punished like this.
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- Jan 2023
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It is thusclear that the biblical author had access to historical information originatingin the 10th and 9th centuries BCE.
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- Aug 2022
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twitter.com twitter.com
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Benjy Renton. (2021, November 16). New data update: Drawing from 23 states reporting data, 5.3% of kids ages 5-11 in these states have received their first dose. Vermont leads these states so far in vaccination rates for this age group—17%. The CDC will begin to report data for this group late this week. Https://t.co/LMJXl6lo6Z [Tweet]. @bhrenton. https://twitter.com/bhrenton/status/1460638150322180098
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- May 2022
-
www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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Embryonal rhabdomyosarcoma (ERMS) of the uterus has recently been shown to frequently harbor DICER1 mutations.
HGNCID:
Tags
- Mutation: c.4420A>G
- Family Information: not identified
- Disease Entity: Embryonal rhabdomyosarcoma (ERMS)
- Mutation: c.5438 A> C
- Mutation: 5428 G>C
- PMID (PubMed ID): 33846547
- Mutation: c.3580delA
- Mutation: c.5125G > A
- Zygosity: Some Cases displayed homozygosity
- case mut: f 28-67
- Mutation: c.5428G>T
- pathogenicity: only 2 of 17 patients died from disease
- Mutation: c.4267G>T
- Mutation: c5113G>A
- GeneName: DICER1
- Inheritance Pattern: Non- inheritance(DNA methylation)
- Variant: Clinvar ID not identified
- case wt: m&f 0.5-19
Annotators
URL
-
- Nov 2021
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www.theguardian.com www.theguardian.com
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Sample, I. (2021, November 2). JCVI failed to back youth Covid jabs despite favourable modelling. The Guardian. https://www.theguardian.com/politics/2021/nov/02/jcvi-failed-back-youth-covid-jabs-despite-favourable-models
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- Aug 2021
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www.themuse.com www.themuse.com
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https://www.themuse.com/advice/the-rule-of-52-and-17-its-random-but-it-ups-your-productivity
I hadn't heard of this before, but it's a thing apparently. Similar to Pomodoro, but it's 52 minutes working and 17 minute breaks instead of 25 and 5.
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- Jun 2021
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outlier-org.admin.datocms.com outlier-org.admin.datocms.com
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Capital Deepening
Start of OS content (late 7.3), runs to the end of the section
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- Mar 2021
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).
AssayResult: 1
AssayResultAssertion: Abnormal
StandardErrorMean: 0
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A total of 84 PALB2 patient-derived missense variants reported in ClinVar, COSMIC, and the PALB2 LOVD database were selected
HGVS: NM_024675.3:c.1653T>A p.(Tyr551Ter)
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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SUPPLEMENTARY DATA
AssayResult: 100.7
AssayResultAssertion: Not reported
PValue: > 0.9999
Comment: Exact values reported in Table S3.
-
To this end, 44 missense variants found in breast cancer patients were identified in the ClinVar database (https://www.ncbi.nlm.nih.gov/clinvar) and/or selected by literature curation based on their frequency of description or amino acid substitution position in the protein (Supplemental Table S1).
HGVS: NM_024675.3:c.2896A>G p.(Ile966Val)
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-
www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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Source Data
AssayResult: 19.46
AssayResultAssertion: Abnormal
ReplicateCount: 2
StandardErrorMean: 1.75
Comment: Exact values reported in “Source Data” file.
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Source Data
AssayResult: 7.03
AssayResultAssertion: Abnormal
ReplicateCount: 2
StandardDeviation: 2.68
StandardErrorMean: 1.9
Comment: Exact values reported in “Source Data” file.
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We, therefore, analyzed the effect of 48 PALB2 VUS (Fig. 2a, blue) and one synthetic missense variant (p.A1025R) (Fig. 2a, purple)29 on PALB2 function in HR.
HGVS: NM_024675.3:c.2006delA p.(E669Gfs)
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www.cell.com www.cell.com
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Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function
AssayResult: 77.5
AssayResultAssertion: Normal
ReplicateCount: 30
StandardErrorMean: 8.6
Comment: This variant had normal function (75-125% of wildtype peak current, <1% late current, no large perturbations to other parameters). These in vitro features are consistent with non-disease causing variants. (Personal communication: A. Glazer)
-
we selected 73 previously unstudied variants: 63 suspected Brugada syndrome variants and 10 suspected benign variants
HGVS: NM_198056.2:c.2291T>A p.(Met764Lys)
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- Feb 2021
-
jmg.bmj.com jmg.bmj.com
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Supplemental material
AssayResult: 80, 99
AssayResultAssertion: Normal
Comment: See Table S3 for details
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Supplemental material
AssayResult: 2.6, 4.8
AssayResultAssertion: Abnormal
Comment: See Table S3 for details
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We analysed a total of 82 blood samples derived from 77 individuals (online supplemental table 3). These 77 individuals corresponded either to new index cases suspected to harbour a pathogenic TP53 variant or to relatives of index cases harbouring TP53 variants.
HGVS: NM_000546.5:c.393_395del p.(Asn131del)
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10.11.66.200 10.11.66.200
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provide information for the following call type ONLY: New Enrollment Card Not Received
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- Jun 2019
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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Statistical analysis
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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T cell proliferation assay
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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Flow cytometry
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Protein extraction and quantification
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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Reagents
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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Central:
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Mesomorphic ectomorph:
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Endomorphic ectomorph:
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Ectomorphic mesomo~ph:
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Endomorphic mesomorph:
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Ectomorphic endomorph:
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Mesomorphic endomor~:
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Endomorph-ectom9££E:
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Mesomorph-endomorph:
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Mesomorph-endomorph:
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Balanced ectomorph:
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Balanced mesomorph:
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Balanced endomorph:
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SOHATOCHART
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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The fluctuation maps (Fu) were calculated from the MD trajectories of the 0-chains as described earlier (Hery et al, 1997). The fluctuation value Fy is given by the equation, where dy(t) is the distance between a pair of designated atoms ( ca atoms as used here) at time t and the angle brackets represent time averages. The Fy values are the standard deviation of interatomic distance. The fluctuation maps in Figure 6 has a black dot wherever the Fu value is less than or equal to 0.5A. Thus dark regions of the map indicate those parts of the molecule which undergo strongly coupled movements.
-
luctuation Maps
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- May 2019
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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Gametocyte rich parasite lysate was prepared using lysis buffer containing phosphatase inhibitors (20IlM sodium fluoride, 20llM ~-glycerophosphate, and IOOIlM sodium vanadate). For some experiments, 2mM calcium or 2 mM EGTA was added to the lysis buffer. IOOllg of lysate protein was incubated with PfCDPK4 anti-sera (1:100 ratio) for 12 h at 4°C on an end-to-end shaker. Subsequently, 50 III of protein A+G-Sepharose (Amersham Biosciences) was added to the antibody-protein complex and incubated on an end-to-end shaker for 2 h. The beads were washed with phosphate-buffer saline three times at 4°C and were resuspended in kinase assay buffer that contained phosphatase inhibitors.
-
mmunoprecipitation of PfCDPK4 from parasite lysates
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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nzyme-linked immunosorbent assay (ELISA) was performed to detect cytokine secretion from human THP-1 macrophages upon activation with LPS. The ELISAs were performed according to manufacturer's protocol. Briefly, THP-1 macrophages were subjected to various treatments and after appropriate time interval the cell culture supernatants were harvested. The capture antibodies for the individual cytokines were diluted 1:250 in coating buffer (0.1 M Sodium carbonate, pH 9.5) and 100 flL was ali quoted into each well of a 96 well ELISA plate (BD biosciences ). The plates were incubated at 4°C for 16 h following which three washes with 0.05% PBS-Tween-20 were given. Blocking was performed using 200 flL of assay diluent (PBS with 10% FCS) per well for 1 h at room temperature following which 1 00 flL of appropriately diluted standards and samples were added and incubated for 2 h at room temperature. A total of 5 washes with 0.05% PBS-Tween-20 were given and the plates were subsequently incubated with 100 flL of detection antibody and streptavidin-HRP for 1 h at room temperature following which 5 washes were given. 100 flL of tetramethylbenzidine (TMB) substrate was aliquoted into each well and incubated for 15 min at room temperature in dark following which 50 flL of stop solution (2N H2S04) was added to terminate the reaction. The absorbance was read at 450 nm and the cytokine levels in the samples were derived based on the OD45o values obtained with standards of known concentration
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Cytokine ELISA
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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with appropriate antibiotics. Serial dilutions of the toxin, made in 0.2% HSA were added to the cells and incubated for indicated time periods. After incubation, the adherent cells were labeled with 0.25 J!Ci eH] leucine per well and the suspension cells with 0.75 J!Ci eH] leucine per well for 3 hat 37 °C. The cells were harvested on to filtermats using an automatic cell harvester, and the incorporation of the eH] leucine in the newly synthesized proteins was estimated using LKB ~-plate counter. Activity was expressed as percentage of control where no toxin was added to the cells. To check the specificity of chimeric toxins for TFR, 10 J.Lg of anti-TFR antibody (HB21) was added to each well prior to the addition of the fusion protein in the competition experiments.
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Cytotoxic activity of restrictocin, its mutants and chimeric toxins was checked on a variety of cell lines. The cellular protein synthesis was assayed in the absence and presence of various concentrations of toxin by measuring eH] leucine incorporation in the newly synthesized proteins. Adherent cell lines namely A431, A549, HeLa, L929 and MCF7 were plated in RPMI 1640 containing 10% FCS at a density of 5x I a:~ cells per well in 96 well culture plates and allowed to adhere for 12 h at 37 °C in the presence of 5% C02• Next day, the medium was replaced with 200 J.LI of leucine free RPMI medium containing 10% serum. The leucine free RPMI containing 1 0% FCS was used for seeding partially adherent cell lines, COL0205 and J774A.1. Cells were allowed to adhere for 12 h and the toxin was added in the same medium. The suspension cell lines, HUT102 and K562 were plated at a density of 5X 103 _cells/well in 80% leucine free RPMI containing 18% complete RPMI and 2% serum immediately before use. The medium used at various stages was supplemented
-
Assay of Cytotoxicity of Chimeric Toxins
-
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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conjugated to fluorescene isothiocyanate ( FITC ) or horse radish peroxidase HRP ) , added. In case of the FITC staining, the cells were finally mounted in a medium containing 0. 1 % para-phenylenediamine PPD and 90 % glycerol in NKH buffer, to retard fading of the fluorescent label. In case of HRP staining, the colour was developed with 0. 05 % DAB ( 3', 3-diaminobenzidine tetrahydrochloride ) in NKH buffer with 0. 002 % hydrogen peroxide. The colour was developed for 15 mins. and the reaction stopped by rinsing in phosphate buffer. The cells were examined under a standard 1 ight microscope for HRP staining or a fluorescent microscope ( for FITC staining ) .
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Cells were grown to subconfluence on polylysine coated glass coverslips contained in plastic dishes. After washing with NKH buffer ( 145 mM NaCl, 5 mM KCl, 15 mM Hepes, pH 7. 4 ) , the cells were fixed for 10 mins. in 70 % ethanol at room temperature. The cells were again rinsed extensively in NKH buffer and the appropriate dilution of primary antibody added. Following OIN incubation at 4°c, the cells were washed 3 X with NKH buffer and the appropriate second antibody,
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Immunocytochemistry.
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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phosphoricacidformedisreducedbytheadditionof1-amino2napthol~4-sulphonicacid(ANSA)reagenttoproducethebluecolor.Theactivityofthebluecolorwasreadat680nmagainstreagentblankusingaU.V.Spectrophotometer.Suitablestandardswererunthrougheachbatchofassays.Theenzymeactivitywasexpressedintermsofpgofinorganicphosphorusformedhr'1mg'1protein.
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Aftereffluentexposure,thecontrolandeffluentexposedfishtissueswereremovedandplacedinabeakercontainingice-coldSEIbuffer(300mMsucrose,200mMNa2EDTA,50mMimidazole,pH7.23)foranalysisofNa+-K+ATPaseactivity.ThetissueswereimmediatelyfrozeninliquidN2andstoredat-80°Cuntilanalyzed. Thespecificactivitiesofsodium,potassium,magnesiumandcalciumdependentATPaseswereassayedaccordingtothemethodsdescribedbyWatsonandBeamish(1980)and Boeseetal.(1982).AdenosinetriphosphatasecatalysestheconversionofATPandADP.Duringthisconversion,onemolecule ofphosphorusisliberated.ATPaseAdenosinetriphosphate^...^Adenosinediphosphate+PTheinorganicphosphorusliberatedwasassayedaccordingtothemethodofFiskeandSubbarow(1925).Inthismethodtheproteinisprecipitatedwithtrichloroaceticacid.Theproteinfreefiltrateistreatedwithaceticacidmolybdatesolutionandthe
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Na+K+ATPase,Mg2+ATPaseandCa2+ATPase
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Thereactionproduction,p-nitrophenolinacidphosphatewasmeasuredspectrophotometricallyat415nmagainstreagentblank.Theenzymeactivitywascalculatedfromthestandardcurveandexpressedasmicromolesofp-nitrophenolformedperhourpermilligramprotein.Therateofhydrolysisofp-nitrophenolphophateisproportionaltotheenzymepresentinthetissue.p-nitrophenylphosphate+NaoH—phosphat?-—>p -nitrophenol+phosphateThecolordevelopedinalkalinephosphataseactivitywasreadat410nmagainstreagentblankspectrophotometrically.Theactivityoftheenzymewasexpressedaspmolphenolformedmin'1mg'1protein
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ThealkalinephosphataseactivitywasestimatedbythemethodofMorton(1955)usingp-nitrophenylphosphatesocolorlessinsolutionbutuponhydrolysis,thephosphategroupliberatesp-nitrophenylwhichishighlycoloredinalkalinesolution
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Alkalinephosphatase(AKP:ortho-phosphoricmonoester-phosphohydrolase;E.C.3.1.3.1)
-
AcidphosphatasewasassayedfollowingtheprocedureofMorton(1955).Theactivityoftheenzymewasexpressedaspmolphenolformedmin'1mg'1protein.
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Acidphosphatase(ACP:acidorthophosphoricphosphohydrolase)(EC3.1.3.2)
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LDHisthekeyenzymeinvolvedinglycolysis,andisresponsiblefortheanaerobicconversionofpyruvicacidtolacticacid,theterminalstageintheEmbden-Meyerhofpathway.Theenzymeactivitywasdeterminedinthecontrolandeffluentexposedfishbrain,gill,muscle,liver,heart,kidneyandair-breathingorgansfollowingSrikantanandKrishnamurthi(1955).TheopticaldensitiesweremeasuredinaUVSpectrophotometerusing340 nmfilterandtheresultsare expressedaspmolesofformazanmg'1proteinhr’1
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Lactatedehydrogenase(LDH)(L-LactateNADoxidoreductase)(EC1.1.1.27)
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Thesupernatant(0.5mlcontaining50mgtissue)wasassayedforSDH.Thereactionwasinitiatedbytheadditionof0.5mlofthesupernatant.Controlsreceived0.5mlsucroseinplaceoftheenzymeextract.Afterincubationfor30minat37°C,thereactionwasstoppedbytheadditionof5mlglacialaceticacidandthederivedformazanwasextractedinto5mloftoluene.Afterkeepingitovernightincold,thecolorwasmeasuredinUV-Spectrophotometerat495mMusingsilicacuvettes.Enzymeactivitieswereexpressedaspmolesofformazanmg'1proteinhr'1
-
Thisisanimportantenzymeinvolvedinthecitricacidcycle.Thehomogenatesofcontrolandeffluentexposedtissueswerepreparedin0.25MicecoldsucroseusingPotterElvehjemtypeglasshomogenizerandcentrifugedat3000rpmfor15min
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Succinicdehydrogenase(SDH)(E.C.1.3.99.1
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Theacetylcholineconcentrationinthetissueswasestimatedspectrophotometricallyat540nmbythemethodofHestrin(1949)usingformicacid-acetonemixture(0.15mformicacidacetone,3:17V/V)astheextractionmediumAchconcentrationwascalculatedintermsofnmolAch/mgtissue
-
Acetylcholine(Ach)
-
Aftereffluentexposure,thetissuesweredissectedout,weighedandhomogenizedin0.25Msucrose.Thehomogenateswerecentrifugedat10,000rev/minatatemperaturebelow8°C.AcetylcholinesteraseactivityofthesampleswasdeterminedatpH7.0usingafinalhomogenateconcentrationof25mgmf1at10°CwithmMacetylcholineiodideassubstrate and0.001or0.002NsodiumhydroxideastitrantfollowingHestron’smethodasgivenbyMetcalf(1951).ProteindeterminationsforalltheChEanalyseswereconductedonaliquotsofthehomogenatesusingamodificationoftheLowryetal.(1951)method.AchEactivityisexpressedinpmolesofacetylcholinechloridehydrolysedmgtissue'1hr'1
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Acetylcholinesterase(AchE
-
0.89%salinesolutioninaTejElon-glasshomogenizerat4°C.Thehomogenatewascentrifugedat4000rpm(3500xg)at4°Cfor20minutes.Theclearsupernatant(organextract)wasusedforestimationofenzymes
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Aftereffluentexposure,thecontrolandexperimentalfisheswerekilledbyhammeringonheadanddissectedimmediately.Excisedbrain,gill,muscle,liver,heart,kidneyandair-breathingorganswereweighed(about20mg)andhomogenizedin2mlof
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Collectionoftissues
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Enzymologicalanalyses
Tags
- Method-17-Method-7-detail
- Method-17-Method-3
- Method-17-Method-2-detail
- Method-17-Method-5-detail
- Method-17-Method-6
- Method-17-Method-3-detail
- Method-17-Method-4
- Method-17-Method-2
- Method-17-Method-1
- method-17
- Method-17-Method-7
- Method-17-Method-1-detail
- Method-17-Method-5
- Method-17-Method-8
- Method-17-Method-8-detail
- Method-17-Method-6-detail
- Method-17-Method-4-detail
Annotators
URL
-
-
shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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The lacZ U118 is an amber nonsense mutation(Am) that confers Lac─phenotype and also polarity of the downstream lacYA genes in the operon due to premature Rho-dependent transcription termination within the untranslated region of lacZ. Melibiose is a sugar which can only be utilized in a lacZ (Am) strain at high temperature (39 ̊C, when the native melibiose permease is inactive) if the downstream gene lacY encoded permease is transcribed and translated. Therefore, in lacZ (Am) strains, growth on minimal melibiose plates (0.2%) at 39°C reflects transcriptional polarity relief at the lac locus, and the same was scored after streaking the relevant strains on such medium
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lacZ (Am) assay
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dependent transcription termination within the untranslated region of trpE. Anthranilate is a precursor of tryptophan, which is the product of trpE-encoded anthranilate synthase. Therefore, in trpE(fs) strains, growth on minimal glucose plates supplemented with anthranilate (100 μg/ml) reflects transcriptional polarity relief at the trp locus, and the same was scored after streaking the relevant strains on such medium
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The trpE9777 is a frameshift (fs) mutationconfers Trp auxotrophy and also polarity on the downstream trpDCBA genes in the operon due to premature Rho-
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trpE(fs) assay
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The galEp3 (galE490∗)mutation represents a 1.3 kb IS2 insertion in the gal leader region (between the promoter and structural genes of the galETKM operon). The mutation causes transcriptional polarity on the structural genes due to rho dependent transcription termination within IS2. In this assay, the gal operon expression in a galEp3mutant or its derivatives was monitored by one of two means. In the first, MacConkey galactose indicator plates (with 1% galactose) were used, where Gal+ colonies are red, and Gal− colonies are white. Therefore, the depth of color serves as an indicator of relative levels of gal expression. In the second method, growth of strains on minimal-galactose (0.2%) was used as a test for Gal+ phenotype
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galEp3 assay
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In vivo transcription termination assays
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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taining withER Tracker™ Blue White DPX: ER Tracker™ is a dapoxyl dye that specifically stains ER in live cells. Since the dye was not found to retain after fixation, live cell staining was performed when fixation was required before staining with antibodies against the CYP proteins. Once washed, the cells were blocked with 3% NGS prepared in 0.001% Digitonin for 30 min at 4°C. This was followed by washing cells with chilled PBS at centrifugation at 805 x g for 10 min at 4°C. Then the cells were incubated with the appropriate primary antibody prepared in 0.001% digitonin for 1h at 4°C. The unbound primary antibody was washed off with chilled PBS three times by centrifugation at 805 x g for 10 min at 4°C. Cells were then incubated with appropriate secondary antibody again prepared in 0.001 % digitonin for 1h on ice. This was followed by three washes with chilled PBS as before. ER Tracker Blue (1p.M) then added to cells which were incubated on ice for 30mins on ice. The excess dye was washed and the cells viewed under the microscope after mounting on slides with anti-fading mounting media
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The protocols for immunostaining cells with different antibodies and dyes were standardized individually for the respective staining procedure: Staining with MitoTracker® Red: MitoTracker® probes diffuse passively across the plasma membrane and accumulate into the active mitochondria. 100]1M stock solutions of MitoTracker Red CMX Ros were prepared in DMSO. Cells were stained at a final concentration of 100nM for 10 min at 37°C, at RT in the dark. The excess unbound dye was washed off and the cells were ready to be viewed under the microscope or processed further for fixation and antibody staining. Staining with antibodies (Table 3.9): Log phase cells were centrifuged at 129 x g for 5 min at RT to remove all dead cells. The live cells were washed 2X with PBS to remove any adherent media and FBS. Fixation was carried out with 2% formaldehyde at RT for 15 min. The fixed cells were washed 2X with PBS by centrifugation at 805 x g at RT for 5 min. They were then blocked using 3% Normal Goat Serum (NGS) prepared in 0.01% Saponin for 30 min at RT. After a wash with PBS, the cells were incubated with appropriate dilution of primary antibody prepared in 0.01% saponin for 1h at RT. The unbound antibody was thoroughly washed off by at least three washes with PBS. Incubation with respective fluorophore conjugated secondary antibody, again diluted in 0.01% saponin, and was carried out for 45 min at RT. The unbound antibody was thoroughly washed off by at least three washes with PBS. The cells were resuspended in a small volume of PBS and mounted on a slide along with anti-fading mounting media (10% glycerol and 0.001% P-phenylenediamine)
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Immunocytochemistry
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between the 5' and 3' flanking regions using the sites Smai and BamHI to generate pBSK+CYP710C1Hyg
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The vector pBlueScript SK+ was used as the backbone to generate the replacement constructs. Standard cloning techniques were used (Sambrook et al., 1989). CYP5122A1 allelic replacement constructs were prepared by inserting ORFs encoding Neomycin or Hygromycin resistance between 0.4 Kb 5' and 0.37 Kb 3' CYP5122A1 flanking regions cloned into the vector pBlueScript SK+, to generate the constructs pBSK+CYP5122A1Neo and pBSK+CYP5122A1Hyg respectively. The following steps were performed. (i) A 416bp 5' flanking sequence of CYP5122A1 was amplified by Hi-fidelity PCR (Table 3.6) and cloned in between unique EcoRV and EcoRI sites of pBSK+ vector. (ii) A 378bp 3' flanking region of CYP5122A1 was amplified using Hi-fidelity PCR and cloned in between BamHI and Xbal sites in pBSK+ with the 5'fragment already cloned in. (iii) ORF for Neomycin resistance was amplified from the vector pXG-GFP+2 and cloned in between the 5' and 3' flanking regions already cloned into pBSK+ vector using the EcoRI and BamHI sites, to generate the construct pBSK+CYP5122A1Neo. (iv) To generate the second replacement construct, hygromycin ORF was amplified from the vector pXG-Hyg (Kindly provided by Dr. Stephen M. Beverley, Washington University) and cloned in between the 5' and 3' flanking regions using the sites Smal and BamHI to generate pBSK+CYP5122A1Hyg. Similarly CYP710C1 allelic replacement constructs were prepared by inserting ORFs encoding Neomycin or Hygromycin resistance between 0.36 Kb 5' and 0.37 Kb 3' CYP710C1 flanking regions cloned into the vector pBlueScript SK+, to generate the constructs pBSK+CYP710C1Neo and pBSK+CYP710C1Hyg respectively. The following steps were performed. (i) A 364 bp 5' flanking sequence of CYP710C1 was amplified by Hi-fidelity PCR (Table 3.7) and cloned in between unique EcoRV and EcoRI sites of pBlueScript SK+ vector. (ii) A 378bp 3 'flanking region of CYP710C1 was amplified using Hi-fidelity PCR and cloned in between BamHI and Xbal sites in pBlueScript SK+ with the 5'fragment already cloned in. (iii) ORF for Neomycin resistance was amplified from the vector pXG-GFP+2 and cloned in between the 5' and 3'flanking region cloned into pBlueScript SK+ vector using the EcoRI and BamHI sites, to generate the construct pBSK+CYP710C1Neo. (iv) To generate the second replacement construct, hygromycin ORF was amplified from the vector pXG-Hyg and cloned in
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Generation of allelic replacement constructs for generation of knock outs
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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A calibration curve of fluorescence intensity values versuspH was prepared for BCECF-AM-loaded wt cells by incubatingcellsin YPD medium containing 50 mM MES, 50 mM HEPES, 50 mM KCl, 50 mM NaCl, 0.2 M ammonium acetate, 10 mM NaN3, 10 mM 2-deoxyglucoseand5 μM carbonyl cyanide m-chlorophenylhydrazone, titrated to five different pH values in the range of 4.0-8.0. Fluorescence intensity values were measured by excitation at 440and 490 nm with emission at 535 nm and a graph was plotted between the ratio of intensity at 490 to 440 nm versuspH. Similar to pHi calibration curve, a polynomial distribution of fluorescent intensity signal and pH was observedfor BCECF-AMprobe
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In vivovacuolar pH calibration curve
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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CgRTT107(3.3 kb),CgRTT109(1.3 kb),CgVPS15(3.4kb) and CgVPS34(2.4kb) ORFs were PCR amplified from genomic DNA of the wild-type strain using high fidelity Platinum Pfx DNA polymerase with primers carrying restriction sites for SmaI-SalI,BamHI-SalI,XmaI-XhoIandSalI-XmaI,respectively.Amplified fragments werecloneddownstream of the PGK1promoterin the pGRB2.2 plasmid. Clones were verified by bacterial colony PCR, sequencing and complementation analysis
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Cloningof C. glabrataORFs
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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Table 2.4: List of double-stranded oligonucleotides used in the present study
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The DNA-protein complexes were then separated from free oligonucleotides on 6.6% native PAGE gel. The samples were loaded into a native PAGE gel, which was pre-run at constant current (40-50 mA) for 15-30 minutes. Electrophoresis was performed at constant current (80-100 mA), till the bromophenol blue dye front reached 1-2 cm from bottom of the gel. The glass plate was carefully removed without disturbing the gel and the Whatmann filter paper no. 3, cut to the size of the gel, and wasplaced over it. The paper was pressed gently and the gel, which was now firmly stuck on the paper, was covered with saran wrap and kept for vacuum drying on the gel-dryer at 80oC for 1 h. After drying, gel was exposed on a Phospho-imager screen for 12-24 h and scanned on Phospho-imager to detect the band of interest.To determine the specificity of the transcription factor binding or sub-unit interacting to the desired oligonucleotide, super-shift assay was performed. For this, 8-10 μg of nuclear extracts were first incubated with desired antibodies (concentration varies for different) or their isotype control for 1h at 25°C, followed by incubation with binding reaction mixture. The various oligonucleotide sequences used in the present study are listed in Table2.4below
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For detection of protein-nucleic acid interaction, an electrophoretic mobility shift assay (EMSA) was conducted as described by Hellman and Fried (2007). Breifly, 8-10 μg of nuclear extract protein was incubated with binding reaction-mixture containing either 32P end-labeled double-stranded oligonucleotides (NF-κB, AP-1, p53 or SP-1) or unlabeled oligonucleotide as shown in table 2.3. The reaction mixture was incubated at 37°C for 45-60 min. After completion of reaction, 6μl of 6X DNA-loading dye was added and mixed well by gentle tapping.Table 2.3: Binding conditions for DNA-protein complexes in EMSA
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Gel shift assay
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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The desired S.cerevisiae strain was grown overnight in YPD liquid medium and yeast cells were harvested by centrifugation at 2,500x gin 15 mL polypropylene tubes. Yeast cells were washed with PBS, suspended in 500 μL lysis buffer (Section 2.1.6.2) and were transferred to a 1.5 mL microcentrifuge tube. Yeast cells were incubated for 15 min on a thermomixer set at 65 ̊C and 750 rpm.After incubation, a volume equivalent to 500 μLof glass beads (0.5 mm) were added and cells were lysedon a beating apparatus by mixing three times for 45 seconds each with intermittent cooling on ice to prevent overheating.Cell lysates were centrifuged at 12,000 x gfor 5 min and upper aqueous phase was transferred carefully to a new 1.5 mLmicrocentrifuge tube, to which 275 μL of 3M sodium acetate was added. To this solution, 500 μL of chloroform was added, mixed well, and centrifuged at 12,000 x gfor 5 min at 4°C (this step was repeated twice). Supernatant was transferred to a new 1.5 mLmicrocentrifuge tubeand500 μL of isopropanolwas added and mixed well by inverting the tube 3-4 times. To precipitate genomic DNA, the suspension was centrifuged at 12000 x gfor 15minat 4°C. Precipitated genomic DNA was washed with 70% ethanol and dried at room temperature. The genomic DNA pellet was dissolved either in 100μLof Sigma molecular biology grade water or TE buffer supplied with Qiagen plasmid mini prep kit, andadd 1 μL of RNase solution (30 mg/mL) was added to this and incubated at 37oC for 1 h. The extracted genomic DNA was checked for integrityon a 0.8%agarose gel by electrophoresis and stored at -20oC
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Genomic DNA isolation by glass bead lysis method
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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44KDa to 670 KDa; Ovalbumin (44KDa), Conalbumin (75KDa), Aldolase (158KDa), Ferritin (440KDa), Thyroglobulin (669KDa)] purchased fromGE Healthcare Life Science. Blue dextran was used for the void volume of the column. The molecular weight of the protein complex fractions was calculated from the plot. The different fractions that were collected as eluates from the column were concentrated,and the presence of WWP2 or WWP1 as a monomer or multimer was identified by western blotting using specific antibodies for WWP2 and WWP1
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The WWP2-WWP1 heterodimeric complex formation under normal conditions and upon cisplatin-induced stress conditions was studied by gel exclusion chromatography. HEK293T cells untreated or treated with cisplatin were harvested,and cell lysate was prepared by using the standard protocol. Sephacryl S-200 (GE Healthcare) columns were equilibrated with 1X NETN (without Triton X-100) at a flow rate of 1ml/min. 0.8ml of cell lysate (1mg/ml) was passed through the Sephacryl S-200 column,and different fractions (fraction size; 500μl) were collected using Bio-Rad 2110 fraction collectorat the same flowrate. To determine the molecular weight of the fractions, column was calibrated with high molecular weight markers [range
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Gel filtration
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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flash (Thermoscientific). β-Glucuronidase activity for GUS was expressed as nanomoles of MU produced/minute/108 cells
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In planta siderophore gene expression was studied by measuring β-glucuronidase activity. GUS marked BXOR1 strain and wild-typeBXOR1 (control) were inoculated in the leaves of 14 day old susceptible Taichung Native 1 (TN-1) variety of rice. After 10 days of inoculation, leaves were crushed and dissolved in 1 ml of MUG extraction buffer without adding MUG substrate (4-methylumbelliferyl β-D-glucuronide). Subsequently, 250 μl extraction buffer containing MUG was added, and incubated at 37°C for appropriate time (Jefferson et al., 1987). Next, 75-μl aliquots were taken from each reaction mixture, and the reaction was terminated by the addition of 675 μl Na2CO3 (0.2 M). Fluorescence was measured against 4-methyl-umbelliferone (MU; Sigma) as standard at excitation/emission wavelength of 365/455 nm, respectively in
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In plantaGUS expression assay for siderophore cluster
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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shRNA were processed for invasion studies. A suspension of 1×105cells in 200μLof 1%FBS containing DMEM was prepared and plated as described in Section 2.2.16. Cells were incubated at 37ºC with 5% CO2for 24 h to allow invasion. Downstream processing of the samples i.e. removing non-invading cells, staining and imaging were followed as described in Section 2.2.16. The number of cells invadedto the lower surface was quantified by counting the total number of DAPI positive nuclei in at least 10 random fields. The total number of cells invaded was normalized to non-targeting control cells and expressed as percentage invasion
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Invasion assays were conducted as described previously (Raoet al., 2015).Invasion chambers with pre-coated matrigel (BioCoat Matrigel invasion chamber, 24 well, 8 μm pore size, Corning) were used to conduct the invasion assays. Packages containing invasion chambers were kept outside and allowed to come to room temperature. Inserts were equilibrated by adding with 500 μLwarm (37ºC) 1% FBS containing DMEM to the upper chamber of the insert and bottom well of the companion plate, and rehydrated for 2 h in a humidified tissue culture incubator at 37ºCwith 5% CO2. The rehydration media was removed carefully without disturbing the pre-coated matrigel matrix on the membrane. Meanwhile, HCT116 cells expressing either non-targeting or IP6K1 targeting
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Invasion assay
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- Dec 2018
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reimaginingthebom.com reimaginingthebom.com
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foolish imaginations of his heart
Constructs which mistake the head as superior to the heart or as an adequate starting place for imaginings will always lead to destruction since the head can not handle nor is it designed for the necessary embedding and recursion which is a seed bearing fruit in itself. But it is sure that, before the brain breaks down and eventually falls/fails, those who mistake it as the best foundation will inevitably turn to mocking them who follow through with flow of spirit through and back to the heart. (see Lehi's Dream 1 Nephi 8:26-27 and Nephi's visitation of the same dream in 1 Nephi 11:35-36)
This phrase,"foolish imaginations" can and ought to be read as the "FULLish imaginations of his heart." In earlier verses we see that a FULL rendering of Lehi's heart brought about fulfillment on several levels already. This "fulfillment" even took the immediate form of a "filament" (pillar of light which struck a rock in front of Lehi just as the communication struck his heart with overpowering energy). The electrical charging effects of Full Imagining can be transmitted beyond the individual and others can be made to feel these effects, however, unless they are allowed to take hold in the heart where embedding and recursion can take place, then they are short lived and sometimes can have disastrous overall effect upon others who rely on second-hand spirit and external motivation. (see 1 Nephi 3:28-31 and 1 Nephi 17:53-54)
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- Sep 2018
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primarydocuments.ca primarydocuments.ca
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MR. CORNELLIER—All the confederations which you have mentioned were or are republican, and had the common fate of republican institutions. You have not said a word about monarchical confederations. MR. JOLY—I have made no mention of monarchical confederations, because none have ever existed, and none can exist. The principle of a monarchy is that the power resides in one person; the principle of confederation is that it resides in all the members of the confederation. A confederation would, therefore, always be a republic, even if formed of several states subject to a monarchy; because the power would not be vested in one person, but in each of the several states, of which no one would acknowledge a head ; it would be a republic consisting of a very small number of members. Before I take leave of all the confederations, the names of which I have mentioned, I intend to say one word, at least, in their favor. We understand that states
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- Aug 2018
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primarydocuments.ca primarydocuments.ca
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The seat of the Federal Government is to be at Ottawa, of course. The Governor General or other head of this magnificent future vice-royalty, or what not, will hold his court and parliament at Ottawa ; but a handsome sop is thrown to Quebec and Toronto, also. They, too, are each to have a provincial court and legislature and governmental departments.
§.17 of the Constitution Act, 1867.
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- Jul 2018
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Local file Local file
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12
Step 17:
Place the desk surface on the studs previously inserted. The two holes should face away from the opening of drawers. Secure the surface by screwing the 1 100218 into the pre-drilled hole using the 100001 tool provided with your package. Consult the graph for proper alignment.
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- May 2018
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transcripts.cnn.com transcripts.cnn.com
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But Theresa May, I think to -- on the heels of the Skripal attack, of the Russian nerve poison on a former agent and for which she garnered considerable international support, riding that wave into this -- and that response was a surprise to her, I think, in many ways, that going into this without getting parliamentary support, which at best would have been ambiguous, given the level of divisiveness and the precariousness of her position in Parliament, meant that she went along with that, without consulting with lawmakers and the backlash has been not just from Jeremy Corbyn or the opposition but also within her party, where there's a lack of consensus on whether or not she should have gone into this with Donald Trump, with French president Emmanuel Macron without greater consultation with the international community.
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transcripts.cnn.com transcripts.cnn.com
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So, you are talking about a soggy weekend, five, six, 700 upwards of 800 millimeters of rainfall in some of this regions across the northern tier of the island of Kawai.
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The attack in Douma was horrific attack on civilians using chemical weapons part of a Civil War that has killed hundreds of thousands of people.
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Again after public outrage grew and the two issues as you say, have come together, one is about the continuing concern about violence against women and people are asking more than five years after that awful rape, gang rape in Delhi in 2012.
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transcripts.cnn.com transcripts.cnn.com
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He ordered a military strike on Syrian chemical weapons facilities but then he hesitated and then reversed direction on advice from his most senior advisers on taking sanctions against companies that helped facilitate that deadly chemical weapons infrastructure by Syria.
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transcripts.cnn.com transcripts.cnn.com
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Well in everything any time that you're talking about Russia and the relationship between the United States and Russia, it's always in the context of this cloud that's over the president about whether or not he's got some kind of ulterior motive for his policies and looked the combination of James Comey going out there over the last several days saying I don't know if the Russians have something that they are using to blackmail the president, and then the sort of abrupt, like hey, I don't want to do sanctions on Russia, you know, yes, there were some sanctions but he was very much pushed in the direction by Capitol Hill and others.
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transcripts.cnn.com transcripts.cnn.com
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$130,000, if that's viewed as a campaign contribution, that is $127,300 above the legal limit.
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transcripts.cnn.com transcripts.cnn.com
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It's $131,000.
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transcripts.cnn.com transcripts.cnn.com
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This was back in 2013 President Obama looked at that chemical attack, and at that point in time, his general was saying, hey, it's not a slam-dunk, because you can detect there were Syrian claims, you can detect Syrians dropped bombs, but it's sometimes hard to know if they dropped an existing vehicle of weapons, and Syria has been known to use chemical weapons, or if the Syrians did it.
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transcripts.cnn.com transcripts.cnn.com
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They are originally from bacteria in fungi and they were designed to be much faster, 10 hundred times faster than the original within a period of five to 10 years.
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transcripts.cnn.com transcripts.cnn.com
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But I'll tell you, some of these flexes that we're having here, one story I picked up that I've never told before or written, but William Webster, former head of the CIA, told me that -- it's one of his best stories, he's allowing me to tell it -- that they created a body double for George Herbert Walker Bush at CIA, meaning somebody who looked just like him and it was uncanny how much CIA had created a human that looked like 41.
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transcripts.cnn.com transcripts.cnn.com
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Do you -- do you -- do you -- I'm sorry, do you disagree with the $4 million for school safety -- $4 billion for school safety, the $6 billion for opioids
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