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Figure 3 with 1 supplement
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() Fluorescence anisotropy of Alexa Fluor594-labeled N130 (at S125C within acidic tract A2; 1 μM) upon titration of the rpL5 peptide; the total [N130] was 200 μM. Insets: light microscopy images of …

Figure 3—figure supplement 1
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SV-AUC profiles for apo N130 (purple), or N130 with rpL5 at 1:1 (light blue), 2:1 (green) and 3:1 (red) stoichiometric ratio (rpL5:N130).

Table 3

Results of sedimentation velocity analytical centrifugation analysis (SV-AUC) of N130 in the absence or presence of increasing concentrations of the rpL5 peptide.

Total concentration in mg/ml.

Sedimentation coefficient taken from the ordinate maximum of each peak in the best-fit distribution at 20 °C with percentage protein amount in parenthesis. Sedimentation coefficient (-value) is a measure of the size and shape of a protein in a solution with a specific density and viscosity at a specific temperature.

Standard sedimentation coefficient (-value) in water at 20 °C.

Molar mass values (MW) taken from the distribution that was transformed to the distribution. The theoretical molar mass is in parenthesis.

Best-fit weight-average frictional ratio values taken from the distribution.

Fit of the data in the 100-300 min sedimentation time frame, for the analysis of the lower MW rpL5:N130 complex intermediates. The MW cutoff of ~ 300 kDa corresponds to a tetramer of N130 pentamers.

Figure 4
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() H-N TROSY-HSQC spectra of apo N130 (magenta), in a soluble complex with rpL5 (2.1:1 rpL5:N130; green) and in the phase separated state (4:1 rpL5:N130; red). () Examples of R and R peak …

Table 4

NMR-derived dissociation constant (K) values determined by monitoring chemical shift perturbations of individual N130 nitrogen backbone resonances while titrating the rpL5 peptide.

Table 5

NMR-derived dynamic parameters for monodisperse apo N130 and a 2.5:1 rpL5:N130 soluble complex.

Dynamic parameters for the N-terminus were determined using a global value; thus, the only residue-specific motional parameter for residues within the N-terminus was (see Materials and Methods).

The second transition in the FA curve (starting at ~2.5:1 rpL5:N130) corresponded to phase separation ( Dolce amp; Gabbana Woman Cotton And Silkblend Macramé Lace Top Black Size 38 Dolce amp; Gabbana Brand New Unisex Cheap Online XB1eVz
). To gain insight into conformational changes associated with phase separation, we performed single-molecule Förster resonance energy transfer ( smFRET) experiments with a mutant of N130, N130 Q15C/S125C , that could be dually labeled with Alexa Fluor594 680 at sites within the pentamer core (Q15C) and the A2 acidic tract (S125C), respectively. The FRET efficiency (E FRET ) for this dye pair within the droplet phase (E FRET ~0.15) was dramatically reduced in comparison with that observed in the absence of rpL5 (E FRET ~0.85; Figure 5 ), consistent with theextension of the A2 tract from the N130 pentamer core due to interactions with rpL5 molecules upon phase separation ( Figure 6a ). In contrast to the monodisperse character of the SANS curves for samples of rpL5 and N130 at 0:1 and 1:1 (rpL5:N130) stoichiometry ( Figure 3b,c ), the curve for the 1:3 sample indicated periodic structural organization within the liquid-like droplets ( Figure 3b , red curve). We interpreted these features in terms of inter-molecular correlation distances (55 Å, 77 Å, and 119 Å; see Analysis methods) due to organization of N130 and rpL5 induced by phase separation. Notably, nascent, higher order structural organization was evident in the SANS curve for a solution of rpL5 and N130 with 2:1 stoichiometry ( Figure 3b , green curve). Assembly of rpL5 and N130 into higher order, soluble intermediates at and above the 2:1 stoichiometric ratio was also demonstrated using sedimentation velocity analytical ultracentrifugation (SV-AUC; Figure 3—figure supplement 1 , Outlet Locations Cheap Price Dress for Women Evening Cocktail Party On Sale White Cotton 2017 10 12 8 Vivienne Westwood Buy Newest Bl3UIV4
). The appearance of high molecular weight species within the detection range (<1 MDa) was accompanied by a progressive decrease in the total mass detected, likely due to sedimentation of rpL5:N130 droplets in the sample cell during the dead-time of the experiment. Above the phase separation threshold (>3:1 rpL5:N130), resonances for residues within the N130 core broadened beyond detection in 2D 1 H- 15 N TROSY spectra but not those for residues within the A2 tract ( Figure 4a ). Chemical shift values indicated that these residues remained disordered.

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"A 19" by László Moholy-Nagy, 1927

July 20, 2016

Posted by

Chelsea Jones

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July 20, 2016 Posted by Chelsea Jones

Hungarian artist László Moholy-Nagy was prolific during his lifetime, seeding a pedagogic spirit that asserted the potential for developing creativity to be ever-experimental. He spenthis life creating a new, modern artistic sensibility and teaching others to do the same. Moholy-Nagy’s enduring influence is carried on through those who draw inspiration from the styles and methods he helped spearhead, particularly in design and photography . Read on to discover more about his life and achievements.

László Moholy-Nagy’s enduring influence is carried on through those who draw inspiration from the styles and methods he helped spearhead, particularly in design and photography

Moholy-Nagy began to produce his first drawings as a respite from war, eventuallysustaining an injury while serving in the Austro-Hungarian army in WWI. After he wasdischarged in 1918,he completed his law degree and began to study art more seriously and crystalizehis modern ideas, joining a group called MA that preached the revolutionary aspects of art. He found inspiration in the Russian Constructivists’ use of art to enact social reform in a rapidly modernizing world, and was also influenced by Dadaism and Suprematism.

He found inspiration in the Russian Constructivists’ use of art to enact social reform in a rapidly modernizing world, and was also influenced by Dadaism and Suprematism.

Expandinginto mediums beyond paintings, he experimented beyondthe camerain his 2018 Newest Holi Vintage doublestrap bikini top Blue Marlies Dekkers Cheap Discounts 3UPKbaK
and photomontage, expanding the potential of photographyto see pastwhat the human eye does . Hisphotograms were created by exposing light-sensitive paper directly to light, creating photographic images without using a camera.

expanding the potential of photographyto see pastwhat the human eye does

After relocatingto Berlin in 1919, hekicked off his teaching career. Moholy-Nagy was appointed master of the Staatliches Bauhaus art school in 1923 by founder Walter Gropius , where he took on the school’s guiding design philosophy and expanded it over his five year period there. Bauhaus incubated a style of design central to the Modernist aesthetic,centered on combining art and architecture in ways that harmonized form withfunction. It was here that Moholy-Nagy best cultivated his avant-garde sensibility and belief that art and technology should be united.

This script allows the user to interact with Postgres in several ways.

It can simply start Postgres:

Or, it can be used to run Postgres and pass parameters to the server:

Lastly, it could also be used to start a totally different tool, such as Bash:

Dockerfile reference for the VOLUME instruction

The VOLUME instruction should be used to expose any database storage area, configuration storage, or files/folders created by your docker container. You are strongly encouraged to use VOLUME for any mutable and/or user-serviceable parts of your image.

Dockerfile reference for the USER instruction

If a service can run without privileges, use USER to change to a non-root user. Start by creating the user and group in the Dockerfile with something like RUN groupadd -r postgres useradd --no-log-init -r -g postgres postgres .

Consider an explicit UID/GID

Users and groups in an image are assigned a non-deterministic UID/GID in that the “next” UID/GID is assigned regardless of image rebuilds. So, if it’s critical, you should assign an explicit UID/GID.

Due to an unresolved bug in the Go archive/tar package’s handling of sparse files, attempting to create a user with a significantly large UID inside a Docker container can lead to disk exhaustion because /var/log/faillog in the container layer is filled with NULL (\0) characters. A workaround is to pass the --no-log-init flag to useradd. The Debian/Ubuntu adduser wrapper does not support this flag.

Avoid installing or using sudo as it has unpredictable TTY and signal-forwarding behavior that can cause problems. If you absolutely need functionality similar to sudo , such as initializing the daemon as root but running it as non- root ), consider using “gosu” .

Lastly, to reduce layers and complexity, avoid switching USER back and forth frequently.

Dockerfile reference for the WORKDIR instruction

For clarity and reliability, you should always use absolute paths for your WORKDIR . Also, you should use WORKDIR instead of proliferating instructions like RUN cd … do-something , which are hard to read, troubleshoot, and maintain.

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An ONBUILD command executes after the current Dockerfile build completes. ONBUILD executes in any child image derived FROM the current image. Think of the ONBUILD command as an instruction the parent Dockerfile gives to the child Dockerfile .

A Docker build executes ONBUILD commands before any command in a child Dockerfile .

If done correctly, I could expect the response to look something like this:

Finally, I had to choose a database to store the profiles. I landed on the Realtime Database from Firebase, because it's a NoSQL database that uses JSON as its storage format. I installed firebase and firebase-admin on npm, then required them at the top of my `app.js` file alongside everything else.

To get writing to and reading from the database working, I had to authenticate Firebase using a specially generated "Service Account" private key. I generated the key in the Service Accounts tab of my Firebase settings, and dropped the corresponding code below the rest of my configurations.

Phew! List recorded: check. App registered: check. File structure set up: check. Database setup: check. Time to actually start using the API.

I took a look at the sample response in the GET users/show endpoint documentation and I determined I’d need to store the user’s name, handle (referred to in the docs as screen_name ), location, description, and profile image URL. I also wanted to save the profile color each user set on Twitter to use as a highlight color for buttons, links, and other accents, so I put the following code inside the client library convenience method I had saved in my `app.js` file.

To prevent situations where differences in capitalization would result in two instances of the same profile (like @julesforrest vs @JulesForrest), I set the handle variable to lowercase.

Then I noticed the profile images being returned were way too small to be useful on a desktop display. By tinkering with the image URL, I found a larger image path that worked better for my needs.

People often have URLs, @handles, #hashtags and email addresses in their profile descriptions, but unfortunately, the Twitter API was returning each description as a simple string. Luckily, I found a tool called Autolinker that would search each string and build out anchor tags in the appropriate places.

To get it working, I installed it via npm, then required it at the top of my app.js file:

Basic usage looked pretty straightforward and it came with a few options out of the box that could be passed as an object to the second parameter, like opening each link in a new window or adding a class to each anchor tag.

However, I wanted to add each user’s custom profile color (saved earlier from the API response) as an inline style on that user’s links, which required writing a custom replacement function. While custom replacement functions can handle some pretty complex configurations, I only tweaked the example from the documentation slightly to add the inline color styles and open each link in a new window. It’s important to note the social network for mentions and hashtags needs to be specified at the top of object parameter to link them properly, which wasn’t immediately clear from the docs.

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