#tle3

[chdarling]

Thank you for writing TLE, I hope you enjoy your time off and take your chance to write some non-fandom stuff, I’m sure it will be amazing!

One small TLE3 question. Are we still getting all of the same POVs in TLE3? That last Snape scene felt like a bit of an ending for him? I guess that’s partly as it was finishing off the book and he will obviously still feature, but was just wondering

We definitely haven’t heard the last of Snape. :)

At this point, I do plan to continue the same POVs through to the end of the story. We may even get a few new ones. Surprises are fun!

Thanks so much for reading!!

[lyrics365]

Paranoia

[Refrain] 7it 7yati ma tel3atch ki bghit bentlii fake A7lam w future zwine kifa9t l3in tle3 kolchi fake Mamsayn ta khir yjini mn ta chi fake hit f3aylhom fake Ghar9 f chak mserfe9 b blaka mktob 3liha koulchi fake [Couplet 2] Stora kay2rkho li fat Li 3chto khlanai paranoia khayf Ta mn nfsi wlat katbanliya khant Mab9it nfr9 bin kdoub ou li fact Sknli fe dmaghi ghi lkhouf Mab9itch nti9 fe dli balia…

[chdarling-tle]

I LOVED the new chapter. Always love James, but this was such a satisfying read - James being confronted so forcefully with an unjust world that he desperately wants to fix, but realizes he can’t. But he CAN do small things. And those small things can make all the difference. Also James interpreting Lily’s inability to look at him bc he’s the sun as anger, just perfect. Just a hunch, and feel free not to answer if this is too spoiler-y, but in your TLE3 first kiss snippet, is the [redacted for TLE2 spoilers] part about training Valmai in quidditch? Feels like something that Lily will learn about and feel so touched by. Continually astounded by you, masterpiece, incredible characterization etc etc etc.

I’m not going to answer about the TLE3 snippet yet, but that is a very popular theory indeed 😌🥰❤️

Thank you so much for this kind message. I’m so pleased you enjoyed the chapter!! I’ve been looking forward to getting to this point in James’s development, so it was a joy to get to share it with you. He’s been progressing through the various stages of ignorance ➡️ denial ➡️ excuses ➡️ despair and now we are at ACTION JAMES! Yay! 😂

Much love to you ❤️❤️❤️

[smartphone-science]

This is part 3 of the bacteria-killing-with-arrows series. The first two parts were bacteria killing each other and bacteria firing powerful spikes. You might want to read these two before delving into this current article.

This current article is based on a study that has just been published and which caused me and my former lab buddy long evenings discussing it and coming up with a great (and fun!) concept.

So I guess you remember the arrow bacteria use to fire into other bacteria. Different lethal toxins are attached to this arrow. When bacteria fire this arrow together with the toxins nearby prey bacteria, this usually leads to death of the prey bacterium.

In this study we look at the very tip of the arrow, because on top of the spike of the arrow sits another protein. We call this protein PAAR and it is a super sharp protein. This sharp needle-like protein helps to punch a hole into the prey bacterium. So this super sharp PAAR protein seems to be quite important for the firing and punching process.

Our beloved bacterium Pseudomonas aeruginosa contains seven different PAAR and ten different spike proteins. However, we still don’t know which PAAR sits on top of which spike and which of the PAARs plays a double role and occupies two spikes.

Here we focused on an arrow that is composed of three G2b spike proteins (purple in the below figure). We already knew that G2b is a special spike protein because it contains an extension which at the same time is a toxin (purple pacman). Then we could also show that the toxin Tle3 (red pacman) binds to the G2b spike and is delivered together with this spike.

This already makes this arrow pretty special, because it carries more toxins than the other arrows that I talked about before. In the previous article I discussed the G4b spike that carries the A toxin, and the G5 spike that carries the B toxin. So in many cases it is one spike – one toxin, while the G2b is one spike – two toxins.

Then we wanted to know which of the many PAARs might binds on top of the G2b spike. We indeed found that when PAAR3 was not present in the bacterium (we also tested a bunch of others!), no G2b and no Tle3 were fired out of the cell. Also, neither Tle3 nor the toxin domain of G2b was delivered into other bacteria. This likely means that PAAR3 (orange) sits on top of the G2b spike.

By accident, we then found that when the PAAR3 is not present in the cell, the A toxin (from the G4b spike) was delivered more efficiently. This was surprising, as this means two things: the A-G4b spike has another PAAR on top than PAAR3. Second, PAAR3 seems to inhibit the delivery of the A-G4b spike.

But that is something new. The G2b-Tle3 spike seems to be preferably delivered than the G4b-A spike. But was that also the case with other spikes? Could PAAR3 together with the G2b spike and its many toxins even be the favoured spike of all arrows?

We tested that by looking at another toxin. We found that the G2a spike with the Tle4 toxin was also delivered a lot more efficient when PAAR3 was missing in the cell.

So this means that there is some kind of hierarchy when it comes to the spikes and arrows to be fired. And it looks as if PAAR3 together with the G2b spike and all those toxins is the master arrow to be fired.

Let’s take a step back and look at the whole picture here.

We know that the PAAR protein is actually the protein that is first binding to the crossbow before the rest of the arrow is loaded. We also know that to each PAAR protein a specific spike (G2b, G4b or G5…) is binding. Thus, it seems as if the PAAR protein is something like the “sorting hat” when it comes to making the decision which of the many spikes and thus which of the partner toxins are delivered. This is actually pretty cool and is shown in the picture below.

So, as I said at the beginning, those pointy PAAR proteins are really important in the whole arrow firing process. Looks as if they are even more important than we thought.

This was the third articles of the bacteria-firing- spikes series and if you want to read more, have a look at those publications or send me an email with questions or drop me a comment. I’m always happy about any type of feedback.

via Science Blogs

[cancersfakianakis1]

Appendix-derived Pseudomyxoma Peritonei (PMP): Molecular Profiling Toward Treatment of a Rare Malignancy

Objectives: Pseudomyxoma peritonei (PMP) is a rare malignancy originating from the appendix, characterized by disseminated mucinous tumor implants on peritoneal surfaces. We examined the role of multiplatform molecular profiling to study biomarker-guided treatment strategies for this rare malignancy. Methods: A total of 54 patients with appendix-derived PMP were included in the study. Tests included one or more of the following: gene sequencing (Sanger or next generation sequencing), protein expression (immunohistochemistry), and gene amplification (C/fluorescent in situ hybridization). Results: Targeted sequencing of 47 genes detected variants in KRAS (81%), GNAS (74%), SMAD4 (16%), and ATM (16%). Mutations were found at low frequencies (n=1 to 2) in APC, BRAF, PIK3CA, MLH1, and TP53. GNAS and KRAS co-occurrence was found in 87%. Protein overexpression was found in epidermal growth factor receptor (83%), cyclooxygenase-2 (73%), cMET (63%), cKIT (58%), and platelet-derived growth factor receptor alpha (58%). Immune checkpoint expression was found in 36% (programmed cell death protein 1) and 18% (programmed death-ligand 1). Surrogate markers of cell proliferation were found at low rates (TLE3 23%, TOP2A 22%), consistent with the slow-growing biology of PMP. Phosophatase and tensin homolog was intact (wild type [100%]) and positive (immunohistochemistry [80%]). Patients exhibited stable microsatellite status and mismatch repair proficiency (93%). Importantly, multidrug resistance protein expression was elevated (100% BCRP, 94% MRP1, 88% PGP). Markers for gemcitabine (RRM1), fluorouracil (TS), oxaliplatin (ERCC1), and irinotecan (TOPO1) chemosensitivities were detected at favorable rates: 93%, 87%, 77% and 65%, respectively. Conclusions: Molecular profiling by multiple platforms identified potential therapies for the nontargetable KRAS-mutated population. The role of cMET-targeted therapeutics and immune checkpoint inhibitors merits further investigation. Biomarker-guided selection of cytotoxic chemotherapies may facilitate efficacy to systemic treatment. https://ift.tt/2A9Dz6K

[chdarling]

Seeing as James has some explicit thoughts keeping him up at night, is there any chance we'll get some explicit scenes in TLE3? Maybe not a full-on nsfw (by the way, would you ever write smut or is that not your thing?) but maybe a soft, poetic lovemaking scene for our beloved jily?

Smut is not my thing but I’ve never denied that there will be sex scenes in TLE :)

[chdarling]

Thank you for the nonstop gift that has been TLE over the last couple years I hope you enjoy your well deserved break <3

My theory for TLE 3 so far is that Petunia was nasty which led to Lily crashing at Sirius’ place which led to her never getting the letter which leads to James thinking she’s ignoring it and wallowing in self pity and ~drama~. Blink once if I’m right

Hehehee no comment 🤭 (and thank you!)

[chdarling]

Florence cameo in tle3 spoiler plz

y’all have no idea how evil I could be if I wanted to

[chdarling]

Heyy CH!! I can't believe TLA2 is almost over and I can't believe how good your writing and the story are 😔 I think I'll reread it in the near future because I already miss it soooo much 💖

So of course I was daydreaming about TLA3 and TLA4 and I have a little question 😁 (No need to answer if you don't want to or think it's a spoiler) TLA1 is almost half the size of TLA2, do you have any idea how big (by chapters or word count) TLA3 and TLA4 will be compared to book 1 and 2? Like, is TLA2 going to be the biggest book among the four or do you imagine that the next ones will be even bigger?

Sending all my love always!!! 💐💐💐

Hahaha look, I’m an unreliable source here. Before TLE2, I swore that it was going to be shorter than TLE1. And then I got to writing it and remembered that 6th year jily is my favorite genre of jily and now the 400,000 word monstrosity that is TLE2 exists. 🙃

So while I fully intend for sjt here and tell you, with a gee whiz gosh and golly earnestness that would put any politician to shame, that TLE3 will absolutely not be a word longer than TLE1, gosh dang it…I’ve been proven to be a dirty liar who can’t be trusted. 🤷🏻‍♀️🤷🏻‍♀️🤷🏻‍♀️

TLE4 gonna be long af though I’m afraid. 🫣

Thank you so much for reading ❤️❤️❤️

[chdarling]

Oooo! I just realized with Lily having no reason to go back to Cokesworth there is absolutely almost zero chance of her running into Snape this summer, unless he makes his way to London.

Which means most likely if he DOES see her, she’ll be with Sirius. Lol

Who knows what will happen!!! 🤪

[chdarling]

are you gonna take a break in between TLE2 and TLE3?

hella

[chdarling]

I can’t believe Sirius told James how Lily felt! You should’ve seen my face as I was reading that bit. Utter shock.

And the angst! James, my dearest James, so reluctant to believe what we all already know. It was so beautifully written, because it really makes sense. James being reluctant, needing to hear Lily say it explicitly after everything. It was beautifully written also because the sense of exasperation from Sirius was evident, not only James’ reluctance. I expect this will bring us some more angst and wonderful things, a crescendo if you will, until that EXQUISITE kiss scene you’ve already published.

I wish we would learn more about how James is feeling regarding Sirius’ move and his Lily revelation. Can’t wait.

Just a quick question before I wrap it up: Should we expect TLE3 to include summer hols scenes? Especially in Sirius’ flat?

And to conclude… I once again must say that I am in awe of your talent and I’m extremely grateful to be able to read your work.

❤️

We will see lots of the summer holiday in tle3, that’s all I’ll say for now :)

And thank you so much! ❤️

[chdarling]

can u please please please please please please make sure lily kisses james some day in front of snape because i would love that BUT NOT LIKE ON PURPOSE like snape juts happens to run into them being all cute thankyou on advance i love your work way too much and am way to attached to your lily is she cried a i cry thankyou for causing me mental illness i still love you

#tle2

Ahahaha

No comment 😂🤐

(And thank you so much)

[chdarling]

Can we have The Alchemy in the playlist for TLE3??

Lolll I was just saying to a friend this song has TLE3 jock james vibes 😂😂

[chdarling]

today I broke my own heart working on TLE and fully ugly cried in the kitchen for a silly amount of time about fictional characters with problems I invented in my own stupid little brain

and then I logged onto the world wide web shortly before bed to see that miss swift released a new music video with aggressive jily vibes IN MY HUMBLE AND CORRECT OPINION

so instead of going to bed I did a little swift-inspired jily doodle to make me feel better about the self-imposed sad

here, pls accept this messy snuggle as an advanced apology

idk how to color yet, I’m gonna learn that next lmao

(James’s glasses fell off btw. It’s fine he’s super not worried about it rn.)