243 Matching Annotations
  1. Mar 2024
    1. Italien hat in den ersten beiden Monaten dieses Jahres 2,9 Milliarden Kubikmeter Erdgas LMG Transporte erhalten Punkt das sind mehr zum ersten Mal mehr als via Pipeline aus Algerien geliefert wurde. Der der Anteil des russischen erdgases fiel von 40% 20 21 auf 4,7% 2023. Die Infrastruktur für die Anlieferung von LNG wird auf fünf große Anlagen ausgebaut. Insgesamt ist der Gasverbrauch vor allem wegen der milden Winter um 10% gesunken. https://www.repubblica.it/economia/2024/03/16/news/gas_gnl_rigassificatori_gasdotti_russia-422324021/

    1. I have the opposite of low hums, I hear incredibly high-pitched sounds. This video contains one of those frequencies. CRT monitors and other tube TVs are incredibly loud to me even if the volume is off.

      same here, tinnitus at 16 KHz. i guess this comes primarily from the power grid, because when i measure my body skin voltage versus power ground, via microphone port of a smartphone and spectrogram app, then i see the noise around 16 KHz. i guess the source are power electronics connected to the power grid, especially IGBT transistors which have a working frequency around 16 KHz. i spent 2000 euros on EMF shielding for my bedroom (YShield HNV80), but that can only shield high frequencies like 2.4 GHz wifi, but not the low frequencies from the power grid - 50 Hz to 16 KHz. i started hearing this after a LSD trip... expanding ones consciousness in a bad setting is a bad idea

  2. Feb 2024
    1. Die großen US-Investmentgesellschaften BlackRock, JPMorgan Chase und State Street haben sich zu großen Teilen aus der Koalition Climate Action 100+ zurückgezogen, in der sich Unternehmen zur Dekarbonisierung verpflichten. Damit werden dieser Koalition 14 Billionen (14.000 Millionen) Dollar entzogen. Republikanische Politiker:innen versuchen schon länger zu verhindern, dass Investionen an Dekarbonisierungszusagen gebunden werden. Die Investment-Gesellschaften argumentieren jetzt, dass Climate Action 100+ nicht nur eine Offenlegung fossiler Investionen, sondern eine Steuerung verlangt habe. https://www.nytimes.com/2024/02/16/business/dealbook/wall-streets-climate-retreat.html

    1. Preisentwicklungen auf dem Ölmarkt werden zunehmend vom algorithmischen Handel beeinflusst, also von Kauf- und Verkaufsempfehlungen von Computern in Verbindung mit dem Handel von Derivaten. Wie relevant die "Commodity Trading Advisors" (CTAs), die meist computergesteuert und mit regelbasierten Strategen arbeiten, bereits sind, ist umstritten. Hintergrundbericht im Handelsblatt. https://www.handelsblatt.com/finanzen/maerkte/devisen-rohstoffe/ctas-die-unheimliche-macht-auf-dem-oelmarkt-und-was-dahintersteckt/100008536.html

  3. Jan 2024
    1. Norwegen erteilt in diesem Jahr 62 Lizenzen für die Exploration von Öl- und Gasfeldern, gegenüber 47 im vergangenen Jahr. Die Steigerung geht auf das Interesse von Öl- und Gasgesellschaften zurück. Gegen den Widerstand von NGOs betreibt Norwegen weiterhin eine Ausweitung der Öl- und Gasproduktion, die zu jahrzehntelanger Förderung führen soll. Stark gewachsen ist dabei das Interesse an der Barents Sea. https://www.reuters.com/business/energy/norway-increases-number-new-oil-gas-drilling-permits-including-arctic-2024-01-16/

  4. Dec 2023
    1. Der Standard fasst Berichte der WMO, des europäischen Wetterdienstes Copernicus und des Österreichischen Alpenvereins zum Schwund der Gletscher 2022 zusammen. Die europäischen Alpengletscher verloren 2022 mehr als 5 Kubikkilometer Eis. Die Schweizer Alpengletscher haben in 20 Jahren ein Drittel ihres Eises eingebüßt. Bei einer Temperaturerhöhung um 2,7° bis 2100 würde allein durch das fast vollständige Abschmelzen der Gletscher in Mitteleuropa, den USA, Westkanada und Neuseeland der Meeresspiegel um 115mm steigen.

      https://www.derstandard.de/story/2000145833249/die-gletscher-schmolzen-2022-im-rekordtempo-besonders-in-europa

      Infografik: https://b.staticfiles.at/elm/static/2023-04-schneeanomalie/index.html?nc9238cb923b983a

  5. Nov 2023
    1. spending a few hours each week in close proximity to a certain kind of teacher changed something about their behavior.

      What kind of teacher do you think has this kind of impact?

    2. eachers convey to their students deep messages — often implicitly or even subliminally — about belonging, connection, ability, and opportunity.

      What does this communication look like?

    3. you can’t clearly identify and measure skills, it’s hard to convince people to take them seriousl

      This is a critique with a priority being placed on data based decisions.

    4. And perhaps what students need more than anything for these positive academic habits to flourish is to spend as much time as possible in environments where they feel a sense of belonging, independence, and growth

      Wow, this seems like it would require a drastic change in school environments.

    5. 16. Assessment

      I wonder what the connection will be between motivation and assessment!

    1. Ein Kommentar in der New York Times weist daraufhin, dass einer der Hauptgründe für das für die geringe Nutzung erneuerbarer Energien die veralteten stromübertragungsnetze sind. In den USA scheitert ihre Erneuerung an den Kosten bzw der mangelnden Bereitschaft, für Sie die Allgemeinheit aufkommen zu lassen Punkt der Artikel erwähnt ähnliche Probleme in Deutschland wo Strom von windkraftwerken im Norden mangelsleitungen nicht nach Süddeutschland transportiert werden kann

  6. Oct 2023
    1. Israel was forbidden to set up sacred stones, pillars: “you shall not set up a pillar (massebah), which the LORD your God hates” (Deuteronomy 16:22).

      Relationship to the first two commandments against worshiping other gods and the use of idols?

      How does this relate to the standing stone found in the room at Khirbet Qeiyafa from the time of David?

      Dates of this text with respect to Khirbet Keiyafa?

    1. Water immobilization is a cool thing! The simplest way to accomplish it is by freezing. But can you think of how water might be immobilized (so to speak) at temperatures above freezing, say at 50°F (10°C)? Think Jell-O and a new process that mimics caviar and you have two methods that nearly stop water in its tracks.

      I learned that science and cooking is always connected. Even if we don't think about it in every day life like when water evaporates or freezes it is chemistry. But what I found most interesting that I learned is how water immobilization works, or to put it more simply the science behind Jell-O. When you add gelatin to water it traps the water molecules in place which creates the sort of liquid and solid hybrid we find with Jell-O.

  7. Sep 2023
    1. https://www.amazon.com/How-Make-Notes-Write-Allosso-ebook/dp/B0B7FSQP35/

      Dan Allosso purchased a 30 drawer card catalog (three sections of 5 x 2 without any base) for $300 in 2022.

      It's pictured on the cover of his book "How to Make Notes and Write".

      Purchased at $10 per drawer.<br /> local sale

      Price mentioned at the end of Dan Allosso Book Club 2023-09-16.

  8. Jan 2023
    1. Recommandation 4. Introduire les notions d’éducation à la sexualité dans les programmes officiels decertaines disciplines concernées, au-delà des disciplines liées aux aspects biologiques et sanitaires et del’enseignement moral et civique.Le cadre actuel pose des difficultés liées l’absence de précisions sur l’organisation des séances d’éducation àla sexualité dans l’article L. 312-16 du code de l’éducation, qui a rendu obligatoires au moins trois séancesannuelles en matière d’éducation à la sexualité en 2001. Les circulaires successives ont défini un cadreopérationnel qui a été modifié à plusieurs reprises. Le cadre actuel décrit par la dernière circulaire, en datedu 12 septembre 2018, n’a pas repris deux points sur les supports horaires et les modalités précises sur laprise en charge concrète des séances qui figuraient dans les circulaires de 1998 et 2003. Les modificationssuivantes − sur le modèle de l’article L. 542-3 du code de l’éduction sur l’organisation de la séance annuelled’information et de sensibilisation sur l’enfance maltraitée95 − permettraient de clarifier ces questionsimportantes sur la mise en œuvre concrète de l’EAS :Recommandation 5. Inscrire au moins trois séances annuelles dédiées dans l’emploi du temps des élèves desécoles, des collèges et des lycées (disposition complétant l’article L. 312-16 du code de l’éducation).Recommandation 6. Attribuer la mission d’organisation des séances annuelles aux chefs d’établissement, enlien avec les comités d’éducation à la santé et la citoyenneté (disposition complétant l’article L. 312-16 ducode de l’éducation).
    2. Recommandation 6. Attribuer la mission d’organisation des séances annuelles aux chefs d’établissement, enlien avec les comités d’éducation à la santé et la citoyenneté (disposition complétant l’article L. 312-16 ducode de l’éducation)
    3. Recommandation 5. Inscrire au moins trois séances annuelles dédiées dans l’emploi du temps des élèves desécoles, des collèges et des lycées (disposition complétant l’article L. 312-16 du code de l’éducation)
    1. Samson, last of the Judges, who kills thelion and obtains honey from its corpse, attaches burning torches to the tailsof foxes, and carries the heavy gates of Gaza upon his shoulders

    Tags

    Annotators

  9. Dec 2022
    1. Cette intuition est juste, puisque c’estégalement une approche par la négative queretient la Convention internationale des droitsde l’enfant (CIDE) pour reconnaître, dans sonarticle 16, le droit au respect de la vie privée :« Nul enfant ne fera l’objet d’immixtionsarbitraires ou illégales dans sa vie privée, safamille, son domicile ou sa correspondance,ni d’atteintes illégales à son honneur ou à saréputation »
    2. Cette intuition est juste, puisque c’estégalement une approche par la négative queretient la Convention internationale des droitsde l’enfant (CIDE) pour reconnaître, dans sonarticle 16, le droit au respect de la vie privée :« Nul enfant ne fera l’objet d’immixtionsarbitraires ou illégales dans sa vie privée, safamille, son domicile ou sa correspondance,ni d’atteintes illégales à son honneur ou à saréputation »
    1. 16 - droit del’enfant à la protection contre les immixtionsarbitraires ou illégales dans sa vie privée, safamille, son domicile, sa correspondance etcontre les atteintes illégales à son honneur et àsa réputation.
    2. Cette intuition est juste, puisque c’estégalement une approche par la négative queretient la Convention internationale des droitsde l’enfant (CIDE) pour reconnaître, dans sonarticle 16, le droit au respect de la vie privée :« Nul enfant ne fera l’objet d’immixtionsarbitraires ou illégales dans sa vie privée, safamille, son domicile ou sa correspondance,ni d’atteintes illégales à son honneur ou à saréputation ».
  10. Nov 2022
    1. The btoa() function takes a JavaScript string as a parameter. In JavaScript strings are represented using the UTF-16 character encoding: in this encoding, strings are represented as a sequence of 16-bit (2 byte) units. Every ASCII character fits into the first byte of one of these units, but many other characters don't. Base64, by design, expects binary data as its input. In terms of JavaScript strings, this means strings in which each character occupies only one byte. So if you pass a string into btoa() containing characters that occupy more than one byte, you will get an error, because this is not considered binary data:
  11. Sep 2022
    1. Tutubox installation guide for iPhone and iPad Method 1 – install Tutubox directly to the device As the first step, you need to clear the cache and history in your Safari browser by navigating to the Safari settings.Now you need to go to the general settings and make sure that there are no certificates available in the certificate list.Then go to Wi-Fi settings and then you need to click configure the proxy. Under the configure proxy you need to paste this URL http://ffapple.com in the automatic field. Turn off Wi-Fi for about 10 seconds and enable it again and check if your proxy is still enabled, if not just go and enable it.The next step is to get the latest Tutubox version to your iDevice. You can use the official Tutubox website (tutubox.io). Visit this website from your Safari browser and then download Tutubx iOS for your device and install it.When the installation process is done, you need to go to Settings, General, Device manager, and then click the certificate and trust it. If you do not do this step, you will get a notification when you are trying to open Tutubox that says the term Untrusted Enterprise Developer.Now you have successfully installed the Tutubox latest version on your iPhone or iPad. You are good to go with installing apps, games, and other tools with Tutubox.

      Get Tutubox iOS 16 for installing the latest tweaks for iPhones and iPad for free

  12. Aug 2022
  13. Mar 2022
  14. Jan 2022
    1. moviepilot.de 5,8/10 IMDB 5,8/10 · 33K · Metascore: 77 Parents Guide

      In den Tiefen des Weltalls, weit entfernt von unserem Sonnensystem, leben Monte (Robert Pattinson) und seine kleine Tochter Willow (Jessie Ross) gemeinsam auf einem ramponierten Raumschiff, dessen Besatzung vor einiger Zeit noch aus vielen verurteilten Schwerverbrechern bestand, die sich mit einer gefährlichen Mission von ihren Strafen freikauften. Mit Experimenten wurden sie von der wahnsinnigen Reproduktionswissenschaftlerin Dibs (Juliette Binoche) gequält, bei denen bis auf Monte und Willow alle ums Leben kamen. Monte ist ein stiller Mann, der sich eine harte Selbstdisziplin auferlegt hat. Doch wenn er mit seiner Tochter zusammen ist, wird aus ihm ein zärtlicher Versorger. Nun sind die beiden die letzten Überlebenden der Crew und nähern sich in völliger Isolation ihrem letzten unausweichlichen Ziel: einem schwarzen Loch und damit auch dem Ende von Zeit und Raum. filmstarts.de

      „High Life“ ist ein schmerzhafter Film, doch es lohnt sich, die Expedition ins Nichts zu begleiten. Wer ein klassisches Weltraum-Epos erleben will, der bleibt besser auf dem Boden. Claire Denis‘ Vision ist kompromisslos und radikal. Ein einzigartiges, schwarzes Juwel. filmstarts.de 4,5/5

      In „High Life“ nimmt uns die französische Arthouse-Regisseurin Claire Denis mit ins Weltall. Von Zivilisation ist da oben aber nichts zu spüren. Stattdessen gibt es eine zwischen Wahnsinn und Klaustrophobie schwankende Stimmung, die nach und nach zwischenmenschliche Abgründe freilegt, während das Publikum vergebens auf Erlösung, Hoffnung oder eine tatsächliche Handlung wartet. Oliver Armknecht 8/10

      An Bord eines Raumschiffes werden übergriffige Experimente durchgeführt, die die Überlebenden vor komplizierte Fragen stellen und dem Publikum schwer zu denken geben. Claire Denis erobert mit ihrem ungewöhnlichen Vertreter eines traditionsreichen Genres reizvolles Neuland. epdFilm ?/10

    1. TMDB (66%) JustWatch (68%) Filmstarts (4/5) Moviepilot (5,4/10) IMDB (5,7/10 · 7,6K | Metascore: 52)

      Handlung

      Ein gemütliches Häuschen auf dem englischen Lande: Der Weihnachtsbaum ist liebevoll geschmückt, das Feiertagsfestmahl opulent vorbereitet und nostalgische Evergreens klingen durch die Räumlichkeit. Als Nell (Keira Knightley), Simon (Matthew Goode) und ihr Sohn Art (Roman Griffin Davis) Verwandtschaft und Freunde in dem kleinen Cottage willkommen heißen, scheint dem perfekten Weihnachtsfest im Kreis der Liebsten rein gar nichts mehr im Wege zu stehen. An der Geschichte gibt es jedoch einen Haken, denn überleben wird niemand die Weihnachtsfeiertage...

      Rezension(en)

      Jüngstes Gericht am Heiligen Abend: Das bitterböse „Was-wäre-wenn“-Kammerspiel von Camille Griffin lässt die Generation „Fridays For Future“ unterm Tannenbaum mit der Dekadenz ihrer Eltern kollidieren – und das ist tatsächlich ein (dunkelschwarzhumoriges) Fest! (filmstarts 4/5)

      „Silent Night“ beginnt wie eine typische Weihnachtskomödie rund um eine dysfunktionale Gruppe, bevor es sich nach und nach in ein Endzeitdrama verwandelt. Die Mischung der einzelnen Bestandteile geht nicht ganz auf, da fehlte dann doch ein schlüssiges Konzept. Sehenswert ist der ungewöhnliche Mix aber, zudem mitreißend gespielt. armknoli

  15. Aug 2021
    1. RRID:ZFIN_ZDB-ALT-040716-16

      DOI: 10.7554/eLife.44431

      Resource: (ZFIN Cat# ZDB-ALT-040716-16,RRID:ZFIN_ZDB-ALT-040716-16)

      Curator: @scibot

      SciCrunch record: RRID:ZFIN_ZDB-ALT-040716-16


      What is this?

  16. Jul 2021
  17. Jun 2021
    1. We explore them in other chapters and in particular in Macroeconomic Policy Around the World.

      Reference to OS 19 (not currently mapped in our AL)--remove?

  18. Mar 2021
    1. 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: 4.7

      AssayResultAssertion: Normal

      StandardErrorMean: 0.11

    2. 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.1600T>G p.(Ser534Ala)

    1. SUPPLEMENTARY DATA

      AssayResult: 102.6

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    2. 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.2865T>A p.(Ser955Arg)

    1. Source Data

      AssayResult: 64.92

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardErrorMean: 8.7

      Comment: Exact values reported in “Source Data” file.

    2. Source Data

      AssayResult: 77.45

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 6.2

      StandardErrorMean: 4.38

      Comment: Exact values reported in “Source Data” file.

    3. 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.1865T>C p.(L622P)

    1. Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function

      AssayResult: 23.2

      AssayResultAssertion: Abnormal

      ReplicateCount: 14

      StandardErrorMean: 7.1

      Comment: This variant had partial loss of function of peak current (10-50% of wildtype) and a >10mV loss of function shift in Vhalf activation, therefore it was considered abnormal (in vitro features consistent with Brugada Syndrome Type 1). (Personal communication: A. Glazer)

    2. we selected 73 previously unstudied variants: 63 suspected Brugada syndrome variants and 10 suspected benign variants

      HGVS: NM_198056.2:c.2254G>A p.(Gly752Arg)

  19. Feb 2021
    1. Supplemental material

      AssayResult: 61

      AssayResultAssertion: Abnormal

      Comment: See Table S3 for details

    2. Supplemental material

      AssayResult: 3.1

      AssayResultAssertion: Abnormal

      Comment: See Table S3 for details

    3. 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.375G>A p.?

  20. Sep 2020
  21. Jun 2020
    1. vous

      Pourquoi est ce que pour cette question il a fallu calculer a nouveau le rayon de la terre alors qu'elle était déjà donnée dans un document et que nous l'avons calculer dans la question 13 ? Et que les 3 valeurs ( valeur donnée dans le doc et dans les calculs ) ne sont pas les mêmes

  22. Jan 2020
    1. RRID:ZFIN_ZDB-ALT-170522-16

      DOI: 10.7554/eLife.42881

      Resource: (ZFIN Cat# ZDB-ALT-170522-16,RRID:ZFIN_ZDB-ALT-170522-16)

      Curator: @evieth

      SciCrunch record: RRID:ZFIN_ZDB-ALT-170522-16


      What is this?

  23. Jun 2019
    1. chains were generated in the same way as for the isolated a-chains. Thus the model consisted of eight polypeptide chains ( 4 a-chains and 4 13-chains) with the central two a and two 13 chains making up the axial contact interface. The simulations of the above fiber models were carried out without explicit solvent using the GROMOS96 vacuum force field as implemented within the GROMACS suite of programs. Models of HbS mutants were generated using the program SCWRL and the mutant fiber models were generated as for the native HbS model. SCWRL replaces only the side chains of desired residues with the best possible rotamer of the mutated amino acids. Thus the initial backbone conformation of the isolated a-chains and the fiber models remained identical in the native HbS and the mutants. The MD simulation protocol for the fiber models consisted of an initial steepest descent energy minimization for 1000 steps followed by full MD simulation for 1.2ns at 300 K. Essential parameters of the simulation like the radius of gyration, root mean squared deviation from the initial structure as well as the kinetic and potential energies are summarized in Chapter I, Table 4. It was observed that all the global indicators of the simulation stabilized to their average values within 0.2ns. Hence data from 0.2ns till the end of the simulations were used for all subsequent analysis
    2. MD simulations on the a-chain of HbS and the mutants were carried out using the GROMACS suite of programs (Lindahl eta!, 2001). The initial structure of the native protein was taken from the high-resolution x-ray crystal· structure (Harrington et a!, 1997) of HbS (PDB entry: 2HBS). The structures of the mutants were generated interactively using INSIGHT-II. The initial model structures were placed in a simulation box of size 42.8 x 31.5 x 42.7 A. The closest distance from any protein atom to the walls of the box was not less than 9 A. The system was then solvated by adding a bath of SPC (Berendson eta!, 1981) waters in such a way that the density of the system was as close to 1 as possible. The overall charge of the system was neutralized by placing suitable counter ions wherever necessary (Chapter 1, Table 4). The resulting system was then energy minimized for 1000 steps using the steepest descent algorithm. This was followed by 0.3ns of position restrained MD during which the solvent and counter ions were allowed to move freely but the protein atoms were harmonically restrained to their initial positions. Finally, normal MD was run for 3ns using the default GROMACS force field. Bond lengths were restrained to their equilibrium values using the LINCS (Hess et a!, 1997) algorithm and a cut-off radius of 0.9nm was used for non-bonded interaction calculation. The temperature of the system was maintained close to 300K by weak coupling to an external temperature bath with a coupling constant of 0.1 ps. The integration time step used throughout the simulation was 1 fs. MD simulations of single mutant a-chains (K 16Q, E23Q and H20Q) as well as the double mutants (K16Q/H20Q, K16Q/E23Q and H20Q/E23Q) were carried out in a similar fashion for 3ns. In order to directly analyze the effects of the mutations on the contact interface, we also carried out MD simulations on a miniature model of the sickle hemoglobin fiber consisting of a complex of two hemoglobin tetramers. In the low salt crystal structure of deoxyhemoglobin S (Harrington et a!, 1997) the asymmetric unit consists of two HbS tetramers that pack as two strands of HbS molecules running parallel to the crystallographic a axis. Axial contacts occur between two tetramers of the same strand and lateral contacts occur between tetramers of different strands. A canonical fiber model was generated by taking one of the two tetramers in the asymmetric unit together with its neighbor translated along the crystallographic a axis [Figure 7 (A, B)]. Fiber models incorporating mutant a
    3. Molecular dynamics (MD) simulatio
  24. May 2019
    1. temperature for 2h. The nitrocellulose membrane was washed extensively with PBST and developed using chemiluminescence substrate from Pierce (USA).
    2. The proteins separated by SDS-PAGE were transferred from the gel to· nitrocellulose membrane using a blotting apparatus (Bio-Rad, USA). In brief, after removal of the stacking gel, the resolving gel was placed over nitrocellulose membrane and sandwiched with Whatman 3 mm filter paper in a cassette. The cassette was submerged in transfer buffer and transfer was carried out at 150 rnA for 3h at 4°C. Following the transfer, the membrane was carefully removed from the blotting apparatus and blocked with 3% non-fat dry milk protein for Ih. The membrane was washed thrice with PBST and incubated overnight with the primary antibody at 4°C. Following incubation, the membrane· was washed thrice with PBST and incubated with appropriate HRP-labeled secondary antibody at room
    3. Western Blot
    1. bacteria. The cells were washed once with ice-cold PBS and the uptake of labeled bacteria was analyzed by flow-cytometry
    2. The phagocytic ability of macrophages was determined by monitoring the uptake of Bioparticles® Alexa fluor 488 labeled dead E. coli (Molecular Probes, Eugene, OR). 2X105 THP-1 macrophages were plated per well in a 24 well plate. Alexa fluor 488 labeled dead E. coli particles were opsonized with an opsonizing reagent obtained from Molecular Probes. These opsonizing reagents are derived from purified rabbit polyclonal IgG antibodies that are specific for E.coli. These opsonized bioparticles® were transferred to the macrophage culture at an multiplicity of infection (MOl) of 1:10, i.e., 10 bacteria per macrophage. The plates were briefly centrifuged at 250 x g to allow the bacteria to settle at the bottom of the plate and were then transferred to an incubator maintained at 37°C and 5% C02 in air for 1 h. The culture medium was aspirated to remove excess unbound bacteria and the cells were washed 3x with ice-cold PBS. To eliminate fluorescence from non-phagocytosed bacteria adhering to the macrophage membrane, 0.25 mg/mL Trypan blue was added and incubated for 10 min to quench the fluorescence of extracellula
    3. Measurement of phagocytic ability of macrophages
    1. Cell-free protein synthesis inhibitory activity of restrictocin was assayed as described by Harlow and Lane ( 1988). The frozen rabbit reticulocyte lysate was thawed on ice in the presence of 30 J..LI of haemin solution per 0.5 ml vial. The toxin was diluted in 0.2% RNase free BSA and several concentrations were incubated with 10 J..LI lysate, I mM ATP, 0.2 mM GTP, 75 mM KCI, 2 mM magnesium acetate, 3 mM glucose, 10 mM Tris-HCI, pH 7.6, 4 J..LM amino acid mixture without leucine, 0.16 J..LCi eH] leucine, 1.33 mg/ml creatine phosphokinase, and 2.66 mg/ml creatine phosphate in a reaction volume of 30 J.LI. The reaction was carried out at 30 °C for one hour and terminated by adding 0.25 ml of I N NaOH containing 0.2% H202• The reaction mixture was further incubated for I 0 min. at 37 °C, BSA added to a final concentration of 65 J..Lg/ml, and the proteins were precipitated with 15% trichloroacetic acid. The mixture was left on ice for 30 min. for complete precipitation and harvested onto 26 mm glass fibre filters. The filter discs were placed in a manifold harvester (millipore) and rinsed with chilled 5% TCA, before the addition of reaction contents. The filters were thoroughly washed with chilled acetone and dried at 37 °C, for one hour. The dried filters were immersed in organic scintillation fluid, and counted using a liquid scintillation counter (Packard).
    2. Assay for Inhibition of in Vitro Protein Synthesis
    1. Following transfection with calcium phosphate or lipofectin, the cells were selected for neomycin resistance by using the analogue G418 Geniticin in the culture medium. After 48 ·hours post -transfection, the cells were harvested and replated at a lower density ( 0.5 x 104 cells I 60 mm dish). Culture medium containing G418 was then added to the cells. G418 was used at two concentrations -400 ug I ml and 500 ug 1 ml. The cells were cultured in G418 containing medium for 2 - 3 weeks. During this period, the mock transfected cells and the cells transfected with plasmid lacking neo gene, died and the transformed cells formed colonies. Individual G418 resistant colonies were picked up and propagated as independent clones. The culture supernates from these clones were analysed by RIA for BhCG. The stability of the BhCG secreting clones was assessed by culturing with several passages over a few weeks in media with or without G418.
    2. Isolation of stable clones.
    1. RFFIT is used for detennination of rabies virus neutralizing antibody (RVNA) titers. It is an in vitro cell culture based technique in which foci of virus-infected cells are observed by fluorescent antibody staining. In brief, mouse neuroblastoma (MNA) cells were cultured in T-25 tissue culture flasks in DMEM supplemented with 10% FCS at 35°C in humidified atmosphere of0.5% C02. For subculturing, cells were trypsinized (0.5% trypsin + 0.2% EDT A in DMEM without FCS), centrifuged at 150 X g for 10 min, resuspended in DMEM supplemented with 10% FCS and aliquoted into T -25 flasks. Sera from immunized mice were heat inactivated at 56°C for 30 min and the RVNA titers were determined by RFFIT as described previously (Smith . et al., 1996). Briefly, 100 111 of various dilutions of the reference (Standard Rabies Immune Globulin, Biological Research and Reviews, FDA, Maryland, US) and the test sera were mixed with 100 111 Challenge Virus Strain-11 of rabies virus (containing 50 FFD50) in 8-well tissue culture chamber slides and incubated at 35°C in presence of 0.5% C02 for 90 min. After the incubation period, 0.2 ml of MNA cells ( 1 x 1 05) were added to each well and the slides incubated for 40 h following which these were fixed in chilled acetone and stained with FITC conjugated anti-rabies MAb (Centocor Inc, USA) for 45 min. The slides were washed three times with PBS, mounted in glycerol : PBS (9 : 1 ), and examined under fluorescence microscope (Optiphot, Nikon, Japan). Data was expressed as neutralizing antibody titer that is the reciprocal of the serum dilution resulting in a 50% reduction in the number of the virus infected cells in the presence of the test serum.
    2. RAPID FLUORESCENT FOCUS INHIBITION TEST (RFFIT)
    1. Immediatelyafterisolation,thetissueswereweighedandsubjectedtolipidextractionthatwascarriedoutinduplicateaccordingtoFolchetal.(1957)
    2. Totallipids
    3. Salineextracts(0.89%)oftissueswerepreparedinTeflonglasshomogenizer.ThecarbohydratecontentoftheextractwasdoneaccordingtothemethodofShibkoetal.(1967)
    4. Carbohydrates
    5. TotalproteincontentwasdeterminedbytheFolin-CiocalteaumethodofLowryetal.(1951)asmodifiedbyZakandCohen(1961).Bovinecrystallinealbuminwasusedasa referencestandard
    6. Totalproteins
    7. Aftereffluentexposure,thecontrolandexperimentalfisheswerekilledbyhammeringonheadanddissectedimmediately.Excisedbrain,liver,muscle,gill,kidneyandair-breathingorgans werepooled incoldcondition andusedforbiochemicalestimations.
    8. BiochemicalAnalysis
    1. Strains were streaked on LBON agar plates and after an overnight incubation at 42°C growth was monitored (compared to that on LBON at 30°C as control). Absence of single colony growth was taken to reflect temperature sensitivity. Whenever needed the phenotype was also quantitatively assessed by plating dilutions of cultures on LBON agar plates and the drop in plating efficiency was scored after overnight incubation at 30°C and 42°C
    2. LBON(Ts) phenotype
    3. This test was therefore used for two purposes: (i) to distinguish relA+ from relA− strains, and (ii) as a qualitative measure of transcriptional polarity relief at the ilv locus. Growth in the presence of amino acids Serine, Methionine, and Glycine (SMG) was scored on glucose-minimal A plates supplemented with each of the amino acids at 100 μg/ml and compared with the growth on non-supplemented glucose-minimal A plates to score for SMG phenotype
    4. The E. coli relA mutants exhibit SMG-sensitive (SMGS) phenotype i.e. growth-inhibition in the presence of Serine, Methionine and Glycine at 1 mM concentration each (Uzan and Danchin, 1978) and is proposed to be a consequence of transcriptional polarity exerted by a frameshift mutation in the ilvG gene on the expression of downstream genes of the ilvGMEDA operon (Lopes et al., 1989). It was observed in another study that the rho and nusG mutants that are defective for transcription termination conferred SMG-resistant (SMGR) phenotype in a relA1 strain (Harinarayanan and Gowrishankar, 2003)
    5. SMG resistance
    6. Lac+ colonies were distinguished from Lac− on MacConkey-lactose plates or on Xgal indicator plates. Xgal is a non-inducing colourless substrate of β-galactosidase enzyme which upon hydrolysis yields dark blue indolyl moieties and hence, the Lac+ colonies on Xgal indicator plates are seen as dark blue colonies. Xgal was prepared as a stock solution of 5 mg/ml in dimethyl formamide and used at a final concentration of 25 μg/ml. On MacConkey-lactose medium (pH around 7.1) on the other hand, Lac+ strains can utilize the lactose sugar present in the medium to lower the pH of the medium to 6.8, resulting in a pink coloured colony while Lac─ strains are unable to utilize lactose to give a white colour
    7. Lac phenotype
    8. Scoring for phenotypes
    1. injection, programmed at 20°C min-1 to 200 °C and held for 10 min, then at 10 °C min-1 to 230 °C, and finally at 5 °C min-1 to 320 °C and held for 5 min. Injection temperature was set at 260 °C. High purity helium was used as carrier gas of 1.0 mL min-1 flow-rate. The spectrophotometers were operated in electron-impact (EI) mode, full scan of 40-550 amu or selected ion monitoring (SIM) was used, the ionization energy was 70 e V. Calibration curve was generated using n-hexane stock solutions of standard ergosterol dilutions (5-300 ng/mL) in duplicates and plotting the peak area versus the concentration (Yang et al., 2009).
    2. Gas chromatography (GC) is a common type of chromatography used in analytic chemistry for separating and analysing compounds that can be vaporised without decomposition, while Mass spectrometry (MS) is an analytical technique that measures the mass-to-charge ratio of charged particles and thus determining masses for determining the elemental composition of a sample or molecule, and for elucidating the chemical structures of molecules. Combined together the technique of GC-MS can be used to identify individual components in a mixture and also quantitate them. The sterol extracts prepared in 3.2.C.14 were dried under nitrogen (N2) gas, resuspended in n-hexane and derivatized with BSTF A (N,O-bis (trimethylsilyl) trifluoroacetamide) containing 1% TMCS (trimethylchlorosilane) at 70 °C for 1 hr. The derivative mixture was dried under N2 (gas) to remove excess BSTFA and subsequently re-dissolved in n-hexane. The column temperature was set at 100°C and held for 5 min for
    3. GC-MS analysis of ergosterol
    4. Leishmania! expression vector pXG-GFP+2 was obtained as a kind gift from Dr. Stephen S. Beverley (Washington University). Full length CYP5122A1 (Ld27) had been amplified by PCR using primers (F27P3/F27P2, Table 3.4) and cloned into pGEM-T Easy vector. The ORF was then excised from pGEMT using Notl and cloned into pXG-GFP+2 vector. The transformants were selected for the insertion of the gene in the correct orientation using restriction digestion. Standard cloning techniques were used (Sambrook et al., 1989).
    5. Generation of pXG-GFP+2-Ld27 (CYP5122Al)
    1. fluorescence by excitation at 440 (pH-independent) and 490 nm (pH-dependent) with emission at 535 nm. Ratio offluorescence intensity at 490 to440 nm was used tocalculatethe vacuolar pH. Background fluorescence was removed by subtracting the fluorescence intensity values of cells without BCECF-AM from the fluorescence intensity values of the probe-loaded cells
    2. Vacuole pH inyeast cells was determined asdescribed previously (Padilla-López and Pearce, 2006). Briefly, log-phase,YPD medium-grown yeast cells were harvested and suspended in 200 μl YPD medium containing 50 μM 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein, acetoxymethyl ester (BCECF-AM; Invitrogen # B1150) to the final cell density of 4 x 107 cells. Cells were incubated at 30 ̊C for 30 min at room temperaturefollowed by three washeswith YPD medium. Washed cells were resuspended in 1 ml YPD medium and 200 μl cell suspension was used for recording
    3. Measurement of vacuole pH
    1. After restriction enzyme digestion, digested products wereresolved on agarose gels and desired DNA fragmentswereextracted from the gel. Concentration of gel-extracted DNA fragments was determined usingspectrophotometerand ligation reactions were set up using a molar ratio of vector to insert of 1:3 and 1:1 for sticky and blunt end ligations, respectively. Ligation mixwas incubatedeither at 22ºC for 4 hor at 16°Cfor 14-16 h. After incubation,T4DNA ligase was inactivatedat 65ºC for 20 min
    2. Ligation
    3. Genomic mapping of disrupted locusin Tn7insertion mutants was carried out as describedpreviously(Kaur et al., 2004).C. glabratamutants carrying Tn7insertionswere grown in YPD-liquid medium and genomic DNA was isolated fromovernight cultures. 10 μg genomic DNA was digested either with restriction enzyme MfeIor SpeI.Restriction enzyme-digestedDNA was precipitated with 1 ml ethanol and 1/10thvolume of sodiumacetate (3 M,pH 5.2). DNA pellet was washed twice with ice-cold 70% ethanol, air driedand was resuspended in sterilewater. DNA was recircularized with T4 DNA ligase.Resultant circular DNA carriedTn7cassette flanked on bothsidesby the disrupted locus oftheC. glabratagenome. CircularDNA wastransformed in E. coliBW23473 strainwhich contains protein Π (the product of the pirgene) required by R6Kγorifor replication.Twoverified transformants were grown overnight in LB-kanamycin medium and plasmids were extracted. Purified plasmids were sequenced withprimers reading outwards (OgRK 183 and OgRK 184) from both ends ofTn7cassette.Sequences obtained were compared,usingBLAST,against C. glabratagenome sequence database and regionsof Tn7insertions in C. glabratawere mapped
    4. Mutant rescue
    1. The mixture was then incubated at 37oC for 45 min to radiolabel the oligonucleotide. Simultaneously, the Sephadex G-50 column was prepared in 1 ml syringe. The reaction mixture was loaded on the column and the eluting fractions were collected in the microfuge tube by loading 200 μl milli-Q water on the top of the column. After collecting 5-6 fractions, the tubes were analysed using a GM counter for the amount of radioactivity. The fractions having specific activity between 3.5-4.5 X 106cpm/pmoles were pooled. To this, 100 pM of the complimentary strand of the oligonucleotide was added and heated at 95°C for 5 minutes. The mixture was allowed to annealat room temperature for 1 h and further used in Gel shiftassay
    2. The oligonucleotides of different transcription factors such as NF-B, AP-1, p53 and SP-1 were 5′-end labelled using radioactive γ32-ATP (obtained from BRIT, BARC, Mumbai, India) and Polynucleotide kinase, as per manufacturers’ protocol. The reaction mixture containing the different components, described in the table 2.2below was added in a microfuge tube.Table 2.2: Various components of oligonucleotide labeling reaction mixture
    3. Radioactive labelling of oligonucleotides
    1. After 14-16 h incubation, hybridization buffer was decanted to a radioactive liquid waste container. Membranes were washed twice with 2X SSC (saline-sodium citrate) containing 0.1% SDS for 15 min at 55°C followed by two washes with 1X SSC containing 0.1% SDS for 15 min at room temperature. Post washes, membranes were rinsed with 1X SSC buffer at room temperature exposed to a phosphorimager screen for 1 h and scanned using a phosphorimager (Fuji Film FLA-9000). The data were analysed by densitometry using Fuji Film Multigauge software V3.11 and graphs were plotted using GraphPad Prism5 software.Note:Depending on signal saturation or non-specificity, high stringency washes were performed starting from 0.5X SSC followed by 0.2X SSC or 0.1X SSC wash buffers containing 0.1% SDSat room temperature
    2. Post-hybridization washes
    1. The reactions were carried out at 30 °C for 15 min in 25μlof ubiquitylation reaction buffer (40mM Tris-HCl at pH 7.6, 2mM DTT, 5mM MgCl2, 0.1M NaCl, 2mM ATP) containing the following components: 100μM ubiquitin, 20nM E1 (UBE1), 100nM UbcH5b (all from Boston Biochem). The bacterially purified MBP-WWP2 and MBP-WWP1 E3 ligases were added to the reaction mixture. The bacterially purified and GST bound GST-protein, GST-p73, and GST-ΔNp73 were used as the substrate in the reaction. After the ubiquitylation reaction, the GST beads werewashed five times with 1X NETN buffer and boiled withan equal volume ofSDS-PAGE loading buffer. The ubiquitination of the substrates was determined by western blotting with the substrate-specific antibody
    2. In vitroubiquitylation assay
    1. grown culture was inoculated in fresh PS medium with or without 50 μM 2, 2’-dipyridyl and grown at 28°C. At regulartime intervals, 1 ml culture was removed to determine OD at 600 nm. Furthermore, for GUS assay, 1 ml culture was centrifuged to obtain the pellet, which was washed once in sterile miliQ water, and resuspended in 250 μl volume of 1 mM MUG (4-methylumbelliferyl β-D-glucuronide) extraction buffer (50 mM sodium dihydrogen phosphate [pH 7.0], 10 mM EDTA, 0.1% Triton X-100, 0.1% sodium lauryl sarcosine, and 10 mM β-mercaptoethanol),and incubated at 37°C (Jefferson et al., 1987). After appropriate time intervals, 75 μl aliquotes were taken from each reaction mixture, and reaction was terminated by adding 675 μl Na2CO3 (0.2 M). Fluorescence was measured against 4-methyl-umbelliferone as the standard at excitation/emission wavelength of 365/455 nm, respectively. Likewise, GFP activity was measured in Varioscan flash (Thermoscientific) at exitation/emission wavelength of 472/512 nm, respectively by taking 200 μl of culture directly
    2. For reporter assay, GUS and GFP marked Xanthomonas oryzaepv. oryzicolastrains and control strains were grown overnight in PS medium. 0.2
    3. Reporter assays with β-Glucuronidase (GUS) and green fluorescent protein (GFP)
    1. and pelleted down at 1200 rpm for 5 min. The supernatant containing 10% FBS and trypsin were discarded and the cell pellet was resuspended in media containing 1% FBS. 5×104cells suspended in 200 μLDMEM (1% FBS) was added to the upper chamber and carefully transferred to the well of the companion plate containing 700 μLof complete DMEM (10% FBS) to serve as chemotactic agent. Inserts were lifted with sterile forceps and placed in the companion plate by avoiding any bubbles at the bottom surface of the insert. Cells were incubated at 37ºC with 5% CO2for 18 h (MEFs) and 24 h (HeLa and HCT116 cells) to allow migration. At the indicated time, cells on the upper surface of the filter were removed carefully by scrubbing with wet cotton swabs and inserts were dunked twice in excess PBS. Cells that migrated to the lower surface of the filter were rinsed twice with PBS, fixed with 4% paraformaldehyde for 15 min at room temperature. Post fixation inserts were rinsed in PBS twice and air dried at room temperature. Filters were cut carefully through the edges using a scalpel blade and placed inverted on a clean slide so that the migrated cells faced the upward direction. Filters were mounted with vectashield DAPI and covered with a clean coverslip. Migrated cells stained with DAPI were counted by imaging multiple fields using anepifluorescence inverted microscope (Olympus lX51,Image-Pro AMS 6.0 acquisition software, 20x 0.45 N.A. objective).The number of cells migrated to the lower surface was quantified by counting the total number of DAPI positive nuclei in at least 10 random fieldsper sample
    2. Transwell migration assays were conducted as described previously (Raoet al., 2015). Transwell inserts (24 well, 8 μm pore size, Costar, Corning)were used to conduct migration assays. Inserts were equilibrated by adding 1% FBS containing media to the upper chamber, as well as lower chamber of the insert (companion plate) and placed in a 37ºC incubator with 5% CO2till use. Cells were harvested, counted using hemocytometer
    3. Transwell migration assay
  25. Apr 2019
  26. Dec 2018
    1. A geographical line has been drawn across the Union, and all the States north of that line have united in the election of a man to the high office of President of the United States, whose opinions and purposes are hostile to slavery. He is to be entrusted with the administration of the common Government, because he has declared that that “Government cannot endure permanently half slave, half free,” and that the public mind must rest in the belief that slavery is in the course of ultimate extinction.

      14 + 16 - The author expresses disdain towards the election of a Republican president, not elected in the South, is considered a hostile to slavery, and plans on abolishing it. These concerns imply that he should not be the president of the South.

  27. Nov 2018
    1. DISOLUCIÓN ANTICIPADA DE LAS CÁMARAS Cuya finalidad es reforzar la mayoría parlamentaria del gobierno a base de una nueva elección electoral. La iniciativa es que el Presidente tenga deliberación con el Consejo de Ministros. Sus aspectos son : 1 Que el decreto este firmado por el Rey y refrendado por el Presidente. art 64 2 Debe de haber una convocatoria electoral. art 68 3 Puede afectar a una o las dos Cámaras. Título III. Sus límites son: -No puede haber una disolución anticipada mediante una moción de censura. art 113. -No puede haber una nueva disolución sin haber pasado un año del anterior. art 115 y art 99 -No puede haber disolución en los supuestos del art.116.

  28. Oct 2018
    1. It is possible that a Federative union of the British North American Provinces would afford to the Canadian Government the – readiest mode of escape from the difficulties and embarrassments which now surround the settlement of the “seat of Government ” question, and I presume that I am right in supposing that, although the ostensible object of the proposed inquiry is the union by Federative bonds with Canada of the other British North American Provinces, the Canadian Government have no less in view the, severance of the bond which now joins the two Canadas in a Legislative Union, and the substitution for that bond of a more elastic tie of a Federal or a Federative character.

      §.16 of the Constitution Act, 1867.

  29. Jul 2018
    1. 11

      Step 15:

      Secure piece assembled in the previous step using the 4 102509 with a Flat-Head screwdriver. Consult the graph for proper alignment.

      Step 16:

      Insert the 8 101345 studs into the pre-drilled holes.

    Tags

    Annotators

  30. Mar 2018
    1. It was desirable the General Government should have the control of the medium through which the trade and commerce of the country was carried on, and that in the establishment of banks, the issue of paper money and in offering to the public the paper representative of their labor, in whatever part of the country, there should be the same legislative security for the people

      §§.91(2)(14)(15)(16) of the Constitution Act, 1867.

  31. Nov 2017
    1. To be safe, always pack all medications in your carry on—never in checked luggage. Airplanes don't have refrigerators. If you have a drug that must be kept cold, ask your pharmacist how to package it safely for travel.

      This is great information for travelers as they often have many questions about where to carry their medications and whether or not they need to be documented in the checked luggage.

    1. Under the ACAA(Section 382.57), the airlines are not allowed to charge you for your dialysis machine.iii

      This is great information to be sharing with clients when educating them about life style changes and considerations.

  32. Jul 2017
  33. Jun 2017
  34. Apr 2017
    1. The committee was moved by the willingness of Canadians to welcome Syrian refugees and the eagerness of Syrian refugees to become contributing members of Canadian society. It was concerned, however, that the Government of Canada is not allocating enough resources to help them integrate
    1. Syrian refugees who receive a 1 rating will be on a list of as many as 10,000 names that the UNHCR office in Jordan will offer to the Canadian government.  (Annie Sakkab for The Globe and Mail) Let’s help our own without forgetting to help the refugees
  35. Dec 2016
  36. Nov 2016
    1. Some blocks in my neighborhood are getting downright spooky – front yards are filling with spider webs and tombstones, and ghosts peek through the bushes. Along with the piles of pumpkins and inevitable candy corn appearing in the supermarket, they are a reminder that Halloween is just around the corner. Americans celebrate Halloween on October 31 by trick-or-treating, displaying jack-o’-lanterns (carved pumpkins) on their porches or windowsills, holding costume parties, and sharing scary stories

      american way of celebrating halloween

  37. Oct 2016
    1. In Wall-E and the Toy Storytrilogy, the pleasure is of the suspen-sion of knowledge—the pleasure of notknowing.
    2. One of the distinct pleasures in Pixar’s films is the pleasure of seeing the deepest of human struggles, timeless philosophical questions projected in and through remote forms of representation.
  38. Aug 2016
  39. Jul 2016
    1. Engage adult new writers with online communities of writers, as contributors, readers, and peers, to foster their self-directed learning, self-study, and persistence.

      I know it was not brought up, but when I have my young students peer review, there is always some tension because people are embarrassed to let their peers read their work? For adult learners, i wonder if that anxiety goes away, or if it is heightened because they are adults and sometimes as adults we like to appear as if we know the correct answers most of the time (or at least this is something I do).

    2. discuss, but do not debate

      This is a great tool but I feel like it is not being modeled enough in society. How can we shift this back into our world before it completely goes by the wayside? Do you think social media has caused a decline in the art of "discussion"?

    3. Know your students

      A lot of these strategies can be interlaced with educating youth as well.

    4. Because adults learn by doing, effective instruction focuses on tasksthat adults can perform, rather than on memorization of content. Be-cause adults are problem-solvers and learn best when the subject is of immediate use, effective instruction involves the learner in solving real-life problems.

      When we conduct PDs, these are good points to know to make our teaching more effective

    5. Because adults need to know whythey are learning something, effective teachers explain their reasons for teaching specific skills

      This is also emphasized at the high school level at times. Students often ask, "Why are we doing this?" "Am I ever going to use this?" Maybe instead of viewing that as whining, maybe it as actually more of an "adult" way of questioning.

    6. Is motivated to learn byinternal, rather than ex-ternal, factors

      I'm not too sure that this is always true. A raise, a degree, or a career choice are technically external factors.

  40. Jun 2016
    1. There are many appealing strengths to the idea that learning should be organized around authentic problems and projects that are frequently encountered in nonschool settings: in John Dewey’s vision, “School should be less about preparation for life and more like life itself.”

      I agree and attempt to incorporate "real life" problems as often as I can into my classroom. The problem I often come across is the textbook's idea of a "real life" problem is often much different from what the students relate with. Still, it always helps to add context, especially in math where the question "where are we ever going to use this?" is asked on a daily basis.

    2. there is an infinite number of numbers between any two rational numbers

      This is a fact that can be really unsettling for students to hear, but is also very important for them to actually think about. It not only helps them understand fractions, but also the concept of infinity. Many students, even in high school just simply do not understand fractions, or are not willing to spend the time to complete problems with fractions.

    3. Gradually, students come to ask self-regulatory questions themselves as the teacher fades out. At the end of each of the problem-solving sessions, students and teacher alternate in characterizing major themes by analyzing what they did and why. The recapitulations highlight the generalizable features of the critical decisions and actions and focus on strategic levels rather than on the specific solutions

      This is challenging for me to get the hang of as a teacher. It is very easy for a class to do an activity, a then have the discussion get taken over by a only a few select students that were faster at catching on to the concepts. You really have to be careful as the teacher to make sure you allow for enough time so that all students are able to finish and form ideas about what they just learned. Then having a system to allow all groups to share something is very helpful.

    4. Tests of transfer that use graduated prompting provide more fine-grained analysis of learning and its effects on transfer than simple one-shot assessments of whether or not transfer occurs

      I like to use this type of graduated prompting when doing discovery based tasks. It allows me to give the appearance of giving students hints, when really I am just trying to get them to remember or realize what they already know.

    5. For example, a group of Orange County homemakers did very well at making supermarket best-buy calculations despite doing poorly on equivalent school-like paper-and-pencil mathematics problems (Lave, 1988). Similarly, some Brazilian street children could perform mathematics when making sales in the street but were unable to answer similar problems presented in a school contex

      This could come back to the idea that students perceive themselves as "bad at math" so in a school setting, they automatically shut down and don't actually think about the problems. But when they are presented outside of an educational context, they don't think about it as math, they just think about it as part of their job or social obligation.

    6. e.g., a person may be performance oriented in mathematics but learning oriented in science and social studies or vice versa).

      I come across this issue often in math classes. I have to be very careful when assigning tasks to students, because it is a very fine line between too easy and too challenging. Especially in math, it seems that there a lot of students who have just accepted that they are "bad at math" so they are quick to give up until someone explicitly shows them the answer and how to do it. This is a big issue that I try to fix with students on a daily basis.

    7. A number of studies converge on the conclusion that transfer is enhanced by helping students see potential transfer implications of what they are learning

      I also like to assign problems where students have to identify the error in another student's work. It seems to help students see common mistakes, and helps prevent them from making them in the future.

    8. Understanding when, where, and why to use new knowledge can be enhanced through the use of “contrasting cases,” a concept from the field of perceptual learning (see, e.g., Gagné and Gibson, 1947; Garner, 1974; Gibson and Gibson, 1955). Appropriately arranged contrasts can help people notice new features that previously escaped their attention and learn which features are relevant or irrelevant to a particular concept

      I like using this in situations where a short-cut may be able to be used for one math problem, but cannot be applied in a similar looking problem. I am always sure to draw attention to the two problems so that students can see the slight differences and why they affect the outcome of the problem.

    9. In the rote method, students were taught to drop a perpendicular and then apply the memorized solution formula.Transfer Both groups performed well on typical problems asking for the area of parallelograms; however, only the understanding group could transfer to novel problems, such as finding the area of the figures below.or distinguishing between solvable and unsolvable problems such asThe response of the “rote” group to novel problems was, “We haven’t had that yet.”

      This is good, and also is the reason I hate teaching my students formulas. I almost always show them and make them practice problems first by figuring out what is going on, rather than giving them a formula. The second I give them a formula, they start blindly plugging numbers in, and cannot complete seemingly simple problems that look slightly different.

    10. The concept of adaptive expertise (Hatano and Inagaki, 1986) provides an important model of successful learning. Adaptive experts are able to approach new situations flexibly and to learn throughout their lifetimes. They not only use what they have learned, they are metacognitive and continually question their current levels of expertise and attempt to move beyond them. They don’t simply attempt to do the same things more efficiently; they attempt to do things better. A major challenge for theories of learning is to understand how particular kinds of learning experiences develop adaptive expertise or “virtuosos.”

      I like this, I know, and try to make it clear to my students, that I am not a complete expert in my field. It is ok for even me to make mistakes, or to admit that I am unsure about something (as long as I eventually find them the answer). When students see this, I believe they become less self-conscious about making mistakes in front of their classmates. It also helps the more advanced students understand that even they still have a lot to learn.

    11. Sometimes, however, students can solve sets of practice problems but fail to conditionalize their knowledge because they know which chapter the problems came from and so automatically use this information to decide which concepts and formulas are relevant. Practice problems that are organized into very structured worksheets can also cause this proble

      My textbooks often do this, but I try to combat it as much as possible by either assigning problems in review sections that are already mixed up, or at least providing practice where problems are all mixed up before a large assessment. I could never understand why books would organize so many problem sets in ways that allowed students to work through them thoughtlessly.

    12. Experts usually mentioned the major principle(s) or law(s) that were applicable to the problem, together with a rationale for why those laws applied to the problem and how one could apply them (Chi et al., 1981). In contrast, competent beginners rarely referred to major principles and laws in physics; instead, they typically described which equations they would use and how those equations would be manipulated

      I often notice this in my classroom. I will teach a concept in a way similar to the expert, where I try to focus on the large concept so that students are able to apply that concept in other situations. However, often times when I hear students explaining what I taught to the other students around them, they will only focus on the single method used to solve that exact problem.

    13. Research shows that students who think that intelligence is a fixed entity are more likely to be performance oriented than learning oriented—they want to look good rather than risk making mistakes while learning. These students are especially likely to bail out when tasks become difficult

      These types of students are always the most challenging for me to teach. I firmly believe that the best way to learn is to try something out, get it wrong, then figure out for yourself why it is wrong and how to fix it. Students that are afraid to be wrong cannot learn in this way until their bad habits are broken.

    14. There is no universal best teaching practice.

      Could not agree more. I have been most successful when I have blended teaching strategies, or have switched strategies based on the goal of the lesson.

    15. This will require active coordination of the curriculum across school years.

      I completely agree that this would be the best environment for students to learn, but it has seemed to me very difficult to actually make it happen. Even within school districts there is often little communication between the elementary, middle, and high schools. And even within each level there is not always a natural flow from one classroom to the next. Having teachers keep classes of students for more than a semester or year at a time would help, but that is generally not the case, especially in core subject areas.

    16. Experts are also able to fluently access relevant knowledge because their understanding of subject matter allows them to quickly identify what is relevant. Hence, their attention is not overtaxed by complex events.

      This is an important skill. So many students seem to think that google can solve all of their problems, therefore they don't have to know/learn as much on their own. But the ability to sift through information and actually determine what is relevant and useful comes from your own understanding of the material.

    17. A common misconception regarding “constructivist” theories of knowing (that existing knowledge is used to build new knowledge) is that teachers should never tell students anything directly but, instead, should always allow them to construct knowledge for themselves.

      It seems to me that there always has to be some material where it is very difficult to draw on prior knowledge as it is completely new to students. On these types of lessons, teaching through discovery can lead to more frustration than good. I love to allow my students to discover concepts on their own, but it simply does not work for everything.

    18. In the most general sense, the contemporary view of learning is that people construct new knowledge and understandings based on what they already know and believe (e.g., Cobb, 1994; Piaget, 1952, 1973a,b, 1977, 1978; Vygotsky, 1962, 1978). A classic children’s book illustrates this point; see Box 1.2. A logical extension of the view that new knowledge must be constructed from existing knowledge is that teachers need to pay attention to the incomplete understandings, the false beliefs, and the naive renditions of concepts that learners bring with them to a given subject.

      Assessing students' prior knowledge is one of the most important tools for a teacher. Being able to relate a difficult concept to a simpler concept that students already understand allows students to come to a much deeper understanding of the content.

    19. At the same time, students often have limited opportunities to understand or make sense of topics because many curricula have emphasized memory rather than under- Page 9 Share Cite Suggested Citation: "1 Learning: From Speculation to Science." National Research Council. How People Learn: Brain, Mind, Experience, and School: Expanded Edition. Washington, DC: The National Academies Press, 2000. doi:10.17226/9853. × Save Cancel standing.

      I see this as a hard line to walk, especially being a math teacher, it is expected for students to memorize certain properties/equations, where as a teacher your focus has to be on getting the students to understand how and when to actually use the equation, and why it works in the first place.

    20. Fundamental understanding about subjects, including how to frame and ask meaningful questions about various subject areas, contributes to individuals’ more basic understanding of principles of learning that can assist them in becoming self-sustaining, lifelong learners.

      I agree, I can generally tell more about what a student knows by the questions they ask, compared to anything else. It is also important as teachers to ask meaningful questions to the students, and avoiding simple yes/no type questions.

  41. Apr 2016
    1. The following autumn I ventured upon a college career against my mother's will.    I had written for her approval, but in her reply I found no encouragement.

      This passage exemplifies the theme of Zitkala Sa's despondent longing for her mother's encouragement and support. Zitkala Sa expresses this theme more as she moves farther away from her mother and continually discriminated against for being different.

  42. Mar 2016
    1. While I thus went on, filled with the thoughts of freedom, and resisting oppression as well as I was able, my life hung daily in suspense, particularly in the surfs I have formerly mentioned, as I could not swim. These are extremely violent throughout the West Indies, and I was ever exposed to their howling rage and devouring fury in all the islands.

      The use of imagery inspires fear and exemplifies Equiano's challenges.

    2. "—No peace is given To us enslav'd, but custody severe; And stripes and arbitrary punishment Inflicted—What peace can we return? But to our power, hostility and hate; Untam'd reluctance, and revenge, though slow, Yet ever plotting how the conqueror least May reap his conquest, and may least rejoice In doing what we most in suffering feel."

      This poem expresses the emotion and sensory experience of perseverance through brutality.

    3. those many instances of oppression, extortion, and cruelty, which I have been a witness to in the West Indies: but, were I to enumerate them all, the catalogue would be tedious and disgusting. The punishments of the slaves on every trifling occasion are so frequent, and so well known, together with the different instruments with which they are tortured, that it cannot any longer afford novelty to recite them; and they are too shocking to yield delight either to the writer or the reader. I shall therefore hereafter only mention such as incidentally befel myself in the course of my adventures.

      Equiano's use of emotion through the times that he is beaten is hard to read. Yet, the way he perseveres through his struggles is captivating.