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wild-aloof-rebel · 4 years

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So since David and Patrick didn’t have the talk we expected this week, what’s your speculation on another natural place it could happen between now and the finale? I’m getting anxious because we’re running out of time for everything 😩

Anonymous said to wild-aloof-rebel: I know you’re prolly getting 100000x asks re: this but are you thrown by tonight? What are your thoughts for the rest of the season’s pacing? I also felt some Open Mic parallels re: the store and its success. Thoughts?

Anonymous said to wild-aloof-rebel: I can’t find that post you made where you basically plotted out the rest of the season, but do you think the wedding will be in the last episode?

Anonymous said to wild-aloof-rebel: if patrick isnt in next weeks episode how will they have enough time to resolve the wedding vs marriage conflict between him and david? at the bachelor party?

Anonymous said to wild-aloof-rebel :since we didn’t get the big come-to-jesus moment about the wedding in this ep, where do you think it’s gonna show up now? i love hearing your thoughts about speculation!

y’all have questions, and i’ve got……… well, not answers, surely, because that would imply that i actually know anything lol, but i’m certainly here to speculate some more.

so my thoughts about the structure of the rest of the season certainly got thrown off a bit with this week’s ep (which to be fair i did say was largely baseless speculation in the first place lol). i didn’t anticipate that they would give us a bit of a breather after the heavy emotions of last week’s ep, but i can appreciate that. i do think from here though we’re gonna jump right back into it. the other story lines aren’t [potentially] quite as cry-worthy [yet], so i do still think we’re gonna kinda tackle the other three roses’ “problems” back-to-back without the need for another breather in between. now though it’s gonna be moira in 6x10, johnny in 6x11, and david in 6x12. nothing’s really changed with my speculation on the moira and johnny fronts, so i’m still running with my original thoughts on those eps being where they have some respective moves to be made. it’s david and patrick’s part of the equation where i had my biggest question mark in the first place, and in that post i put their part as happening in 6x9 to make that nicely rounded four eps for four roses thing happen, which obviously didn’t happen. and so now i feel like 6x12 is quite possibly where that discussion is going to be.

there are a few reasons for that. for one, i’m still not entirely certain that patrick is going to be in the coming ep, david for sure is going to visit a caterer, but i think it’s def possible that patrick isn’t there for it. (my guess rn is actually that david goes with stevie and/or johnny.) i know that some people have wondered if the mention of “personal dilemmas” in the description for 6x11 means that they have their discussion there, but based on what we’ve seen of them in the trailer, i actually think that they’re largely the ones trying to keep everyone else from falling apart in that one. everything that they say and do in those scenes seems really over-the-top, like they’re really laying it on thick that we’re all having fun and everything is fine! they seem pretty united in that effort from the bit we’ve seen, so i don’t get the impression that their disagreement or discussion or whatever you want to call it is going to happen there–i think they’re too busy dealing with everyone else’s problems to tackle their own. but 6x12… the description that we now have for it gives me a very dangling on a cliff edge kind of vibe–like a held breath. there’s a tension to it, with the family seemingly awaiting their fate in some way, and that kind of already-tense setting would be an easy place for an argument to crop up. emotions are already running high as they await news, and, especially if that news revolves around the future of the roses in sc, it would be a really logical place for a discussion of david and patrick’s future to fit in.

i do think, in that case, that that probably pushes the wedding into the finale. it’s possible it’s 6x13, but their arc this season really is largely about the wedding (and even the season promos are about the wedding), and so at the end of the day it probably just makes the most sense for the wedding to be in the finale. i keep trying to convince myself otherwise mostly just because i want it to be earlier so we get some time with them as husbands, but from a writing perspective, yeah, it’s probably gonna be the finale. otherwise their arc for the season is really over before we get to the end, and then whatever else they’re doing–no matter how much we might enjoy just watching them eat breakfast together or whatever–doesn’t really hold any weight in the story.

either way, at this point i think i’m just gonna assume it’s the finale, and then that means either i’m right about it or i’m pleasantly surprised when i’m wrong lol. that just seems like the most optimistic way to go about things.

#anon #replies #speculation #s6 #s6 spoilers #i still am not quite sure how i became the go-to person to ask about speculation #but here we are #not upset about it--just always touchingly baffled lol #but you asked for my thoughts so here they are #i forgot to answer the open mic part of that one anon #but YES big open mic vibes for the RA bits #bea and i talk about that in this week's podcast too!

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unfolded73 · 4 years

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My obligatory post about this blog and Schitt’s Creek Season 6:

I’m going to drop offline entirely from 8:00-9:30pm EST Tuesdays (7:00-8:30pm local time). While I have gotten access to CBC Gem and will occasionally use it to watch an hour early (like when we’re in line at Noah’s show on the night of 6x10, goddammit Noah why), most of the time I won’t be doing that in order to preserve family harmony. Most weeks I expect I’ll be watching at 8pm local time.

I’ll tag “sc s6″ for everything all season and “sc spoilers” for at least a week after something has aired. Block those tags as you need to.

#.#..#...#....#.....#psa #schitt's creek

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fiona-widdershins · 2 years

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Hi Persy. It’s your SC valentine!

Just hoping to find out what your top episodes are from the show?

💝💝💝

Hey there!! Sorry for not replying sooner, covid has been kicking my ass. My top episodes, in no particular order (expect for number one), are:

6x13

3x13

5x11

6x07

5x13

5x14

6x10

6x14

6x05

4x13

#Answers to Things #Anonymous #*scvalentine

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muabannhadatquan7 · 4 years

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Nhà 2 Lầu 180m2, HXH 803 Huỳnh Tấn Phát, Quận 7

NHÀ 2 LẦU HXH 803 HUỲNH TẤN PHÁT, P. PHÚ THUẬN, QUẬN 7 _______________________________ - Diện tích: 6x10, hướng Nam - Kết cấu: Trệt, 2 lầu đẹp, 4PN, 3WC - Gần khu BT Tấn Trường, CC Ngọc Lan, Coopmart, trường học, chợ,.... - Vị trí: Cách ngã tư Huỳnh Tấn Phát giao Nguyễn Thị Thập 200m, cách bệnh viện Quận 7, siêu thị BigC chỉ 5 phút. - Tiện ích: Khu vực đầy đủ tiện ích: Các TTTM lớn như SC Vivo City, Crescent Mall,.. Bệnh viện, các trường quốc tế Canada, Nam Sài Gòn,…. - Pháp lý: Sổ hồng riêng, khu dân cư hiện hữu, hoàn công đầy đủ. ===================== Giá bán: 4.1 TỶ ( Thương Lương ) Liên hệ: 0963.080.823. Triệu

#MuabannhadatQuan7 #NhadatCafeland source https://nhadat.cafeland.vn/nha-2-lau-180m2-hxh-803-huynh-tan-phat-quan-7-1331047.html

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cherylsblossom · 7 years

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yeah a part of me wishes the show ended at 610 but the drawback of that is that scott didn't have a huge role in that season and it would feel kind of wrong if it ended there. the sciles ending would've been perfect and i would've cried though. i guess we can't have it all. and i totally agree on the sc@lia point. in this house sc/ira is endgame -ts anon

in this house we’re still crying about sc/allison but would’ve been totally happy w/ a sc/ira endgame bc they were in love and they were cute and he remembered her name

part of me wishes 6a had been a few more episodes so we could’ve had a sciles plot going on as well (watching scott feel like something was missing and trying to rediscover his best friend) and then we could’ve ended the show w/ the original squad graduating and sciles driving off together

whatever the last scene of the finale is there’s no way it’s better than the mcmartinki locker scene and sciles driving away in the jeep

i’m comforting myself about the apparent lack of stydia in the finale by saying that no other ship got what we did in 6x09 and 6x10 where lydia’s love for him literally ripped a hole into another dimension. truly iconic. name a better ship reunion than the locker room kiss 2.0. name more iconic lines than ‘i didn’t say it back’ ‘you don’t have to’.

#ask #anon #stydiapositivityproject #this bitch is trying to remain positive in the face of the writers' lack of talent

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noahreids · 3 years

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Schitt's Creek Season 6 ⟲ Redux | 6x10 (2/2)

#schittscreekedit #schittscreekgifs #bbsc creations #bbelcher #chewieblog #tvgifs #alexisroseedit #moiraroseedit #schitt's creek #alexis rose #moira rose #my sc edit #my edit #sc: s6 #sc: sunrise sunset #sc: 6x10 #s6redux

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zalrb · 7 years

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do u think elena was a little jealous seeing sc hug in 6x10?

nah i just thought she was sad because liz was dying

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serieslc-blog · 6 years

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Arrow 6x10

Arrow 6×10

[sc name=”LATINO OPENLOAD” lINK=”https://serieslc.net/arrow/” opcion2=”https://openload.co/embed/stPDbjZTLz0/%40rr%40w_S06E10_Lat.mkv” ]

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bulletforged · 7 years

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teen wolf + the hunger games :-)

SEND ME A FANDOM // ACCEPTING.

TEEN WOLF:

the first character i ever fell in love with: chris argent :’)

a character that i used to love/like, but now do not: i’m not sure.

a ship that i used to love/like, but now do not: again, not sure. i pretty much still like all the same ones and dislike all the same ones as i used to.

my ultimate favorite character™: chris argent my precious son i love him so much.

prettiest character: to me, chris. but also allison she’s beautiful and precious and i love her but also lydia and also scott precious beautiful son and melissa is so beautiful erhbgjfnsdm i love them all so much don’t make me pick one they’re all pretty okay 90% of this cast is stunning.

my most hated character: uhhhhh gerard argent :/

my OTP: mcarGENT we started as a little pedal boat and now we have a respectable sized ferry i’m so proud it’s been my ship since s1 and we’re officially sailing.

my NOTP: pet*pher keep it far away from me and ic chris thanks

favorite episode: i mean any episode with a chris + allison cuddle gets me tbh. all of season 3 is so good for them. but also 6x10 cause that mcargent kiss literally made my year bearable.

saddest death: i’d say allison but i mean it didn’t actually happen and she’s still alive and canon got it wrong so??? bye jeff.

favorite season: season 3a

least favorite season: season 5

character that everyone else in the fandom loves, but i hate don’t love: stiles. now there are people on this dash who make me like him again but honestly the majority of his fandom/the fandom in general makes me dislike him.

my ‘you’re piece of trash, but you’re still a fave’ fave: kate argent tbh.

my ‘beautiful cinnamon roll who deserves better than this’ fave: chris deserves better and melissa deserves better and scott deserves better and malia does and lydia does and yeah all of the people in this show deserve better.

my ‘this ship is wrong, nasty, and makes me want to cleanse my soul, but i still love it’ ship: i lowkey ship a really fucked up toxic hate thing with chris and jennifer blake lmao

my ‘they’re kind of cute, and i lowkey ship them, but i’m not too invested’ ship’: st*dia cause i’m team sc*dia all the way tbh.

THG:

the first character i ever fell in love with: haymitch abernathy

a character that i used to love/like, but now do not: not sure

a ship that i used to love/like, but now do not: hayffie oops

my ultimate favorite character™: haymitch or johanna mason.

prettiest character: buttercup.

my most hated character: katniss’ mother. :/

my OTP: um my old otp is in the ‘used to love but now don’t’ category so i’m lost here. but um... haymitch/hazelle hawthorne will always have a special place in my heart.

my NOTP: haymitch/katniss like pls he is her dad can we not.

favorite episode: n/a

saddest death: mags legit rip my heart out and let me eat it cause it’d hurt less than mags’ death. also prim’s. wtf @ suzanne collins. why. and the movie did finnick’s death beautifully tbh that hurt too.

favorite season: fave book/movie is catching fire.

least favorite season: mockingjay part 1. and book = mockingjay too.

character that everyone else in the fandom loves, but i hate: uhh finnick is overrated but i don’t hate him. i just don’t love him.

my ‘you’re piece of trash, but you’re still a fave’ fave: effie trinket and president coin.

my ‘beautiful cinnamon roll who deserves better than this’ fave: peeta mellark i mean come on.

my ‘this ship is wrong, nasty, and makes me want to cleanse my soul, but i still love it’ ship: n/a.

my ‘they’re kind of cute, and i lowkey ship them, but i’m not too invested’ ship’: like every ship in the series tbh.

#CATEGORY // ANSWERED.#thanks for sending these in!#also squinty eyes @ you cause thg is my old fandom and idk who knows that #girlterrors

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noahreids · 3 years

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2020 Schitt’s Creek GIF Advent Calendar ↳ Day 8 | A character who only appears in one episode: Clifton Sparks

#schittscreekedit #schittscreekgifs #schitt's creek #sc advent #bbsc creations #my edit #my sc edit #moira rose #clifton sparks #MUFFY!#sc: s6 #sc: 6x10 #sc: sunrise sunset

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ourhaileydavies · 5 years

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Sequential Delivery of Growth Factors from Hydrolytically Degradable Silica-Based Nanoparticles for Cartilage Tissue Engineering-Juniper publishers

Abstract

Articular cartilage shows a very limited self-healing capability due to it's a vascular structure. It has been reported that sequential supplementation of various growth factors such as bone morphogenic protein 7 (BMP-7), transforming growth factor-beta 1 (TGF-01) and insulin-like growth factor-I (IGF-I) play critical roles in reconstruction of articular cartilage tissue. The objective of this work is to design a drug delivery system which facilitates the controlled sequential delivery growth factors to maximize the extent of chondrogenic differentiation of mesenchymal stem cells and consequently reconstruction of damaged cartilage tissue. Surface-modified silica nanoaprticles (SNs) with short segments of lactide (L) and glycolide (G) units were used for grafting and timed-release of growth factors. It was shown that by changing the length of L and G units, the release rate of grafted bovine serum albumin (BSA) on the surface of SNs can be controlled. Nanoparticles with bigger G units showed faster release rate of BSA compared with shorter G units. Additionally, the presence of short LG segments did not significantly change the size distribution of SNs.

Introduction

Articular cartilage can resist a significant amount of mechanical stress and provide a lubricating surface for the gliding joints and a load-bearing matrix attached to the underlying bone [1] . However, due to its avascular nature and low metabolism, it has a very limited self-repair capability upon suffering a trauma [2] . The disease control and prevention center reported that nearly 27 million Americans suffer from joint pain and stiffness, loss of function and disability [3]. While various strategies such as transplantation of autogenous or allogenous chondrocytes, or the use of mesenchymal stem cells (MSCs) are currently used for cartilage treatment, these strategies always suffer from inherent risks of an immune reaction, lack of suitable donor site, and more importantly they rarely restore the full function to the joint [4,5].

It has been demonstrated that different types of growth factors such as bone morphogenic protein 7 (BMP-7), transforming growth factor-beta 1 (TGF-β1) and insulin-like growth factor-I (IGF-I) play critical roles in tissue engineering of articular cartilage to induce chondrogenic differentiation of Mesenchymal Stem Cells (MSCs) [6-8]. However, the bioavailability and bioactivity of these growth factors are both time and concentration dependent [4,9,10]. Several studies have shown that the sequential supplementation of growth these factors is critical to prevent dedifferentiation of cells by first promoting proliferation with one specific growth factor, and then differentiation and expression of a desired phenotype with another [9,10]. A critical barrier to progress in this way is the lack of suitable delivery systems which work for precise controlled and orchestrated time-dependent delivery of multiple growth factors, although many studies have been conducted to control the delivery of growth factors for different tissue engineering applications. Accordingly, the main challenge of this work is to engineer a programmable delivery system to control the release rate of BMP-7, TGF-β1 and IGF-I growth factors and consequently maximize the chondrogenic differentiation of MSCs.

To address this challenge, the idea is to engineer a silica based nanoparticle system containing short segments of biodegradable polymers such as polylactic acid and polyglycolic acid to control the release rate of grated proteins (Figure 1). We have chosen MSNs given their demonstrated biocompatibility, osteogenic potential[11] , and efficacy as drug delivery vehicles for sustained release of antibiotics [12] and anti-cancer drugs [13]. We envision that due to the hydrophilic nature of silica nanoparticles, bioactivity of the grafted proteins will be significantly enhanced over solid hydrophobic micro/nanoparticles such as poly lactic-co- glycolic acid (PLGA) which are currently used for drug delivery applications. Accordingly, we assume that the protein release rate can be tuned by type or length of the degradable segments. The novelty of this project is to design a low-cost hydrolytically degradable nanocarrier system which facilitates a programmable sequential delivery of multiple growth factors with a controlled timed-release to enhance chondrogenic differentiation of MSCs for cartilage tissue engineering applications.

Materials and Methods

Materials: Lactide (L) and glycolide (G) monomers with >99.5% purity (Ortec, Easley, SC) were dried under vacuum at 40°C for at least 12h before use. N, N/-disuccinimidyl carbonate (DSC) and bovine serum albumin (BSA) were received from Novabiochem (EMD Biosciences, San Diego, CA) and Jackson Immuno Research (West Grove, PA), respectively. Hydrophilic silica nanoparticles (SN) was kindly donated by Evonik Corporation (New Jersy). All other reagents were purchased from Sigma Aldrich (St. Louis, MO).

Methods: To activate the silanol groups of SNs were activated by adding 1.5x10-4 mol triethyl amine (TEA) to 250mg of SN containing 3x10-4 mol SiOH groups in 70ml toluene as the solvent. In the next step, 6x10-4 mol of isopropyl alcohol was added to the mixture to continue the reaction for 2h at 50 °C. Then, the desired amount of L was added to the mixture along with 1ml tin (II) 2-ethylhexanoate as the reaction catalyst and the reaction was run for 6h. The lactide chain-extended silica was used as an initiator for chain extension with G monomer with a predetermined L to G ratio. The reaction was allowed to proceed for 6h at 50°C and the product was precipitated in ice-cold hexane to remove the unreacted monomers. In separate reactions, the mole ratio of L and G was changed from 100% (L100) to 75% (L75/G25) and 50% (L50/G50) while the total amount of L and G was kept fixed at 3x10-3 mol. The synthesized copolymer on SNs surface was functionalized with succinimide groups by reacting hydroxyl end-groups of the copolymer with DSC as we described previously [14]. The product was purified by dialysis against DI water and lyophilized.

To attach BSA on the surface of modified SNs, 10mg SNs was suspended in 0.5mL PBS by sonication for 1 min. Next, 0.5mL of the protein in PBS (20mg/mL for BSA) was added to the SNs suspension. The amine group of the protein was allowed to react with succinimide end-groups of LG in the surface of SNs under ambient conditions for 12h as we previously described (Figure 1) [15]. The protein grafted SNs were freeze-dried to obtain a free-flowing powder. To determine grafting efficiency, the protein grafted SNs were resuspended in PBS and centrifuged at 18,000 rcf for 10min and the supernatant was analyzed for total protein content with the ninhydrin reagent as we described previously [16]. Grafting efficiency was determined by dividing the amount of attached protein (total - free protein) by the initial amount in the grafting reaction.

Size distribution of the SNs was measured by dynamic light scattering with a Submicron Particle Sizer (Model 370, NICOMP, Santa Barbara, CA) as described previously [16,17]. For measurement of release kinetic, 1mg protein grafted SNs were incubated in 1mL PBS at 37°C as we previously described [15]. At each time point, the suspension was centrifuged at 18350 rcf for 10min, the supernatant was removed, the SNs were resuspended in 1mL fresh PBS and incubated until the next time point. The amount of BSA in the supernatant was measured with the ninhydrin reagent as described [16,18].

Results and Discussion

The calculated grafting efficiency of BSA to SNs based on the procedure explained in the method section was 52±9%. The effect of L and G segments on average diameter and size distribution of nanoparticles are shown in (Figure 2a). It can be observed the addition of short L and G parts in SNs does not significantly change the average diameter of particles. The particle sizes are between 120±12 and 165±15 for SN and SN- L100, respectively. Figure 2b reveals the effect of the length of L and G segments on release rate of BSA from nanoparticles. The results demonstrated that by increasing the length of lactide the release rate of BSA will decrease significantly. The cumulative release percentage of BSA is 51±5, 65±8 and 91±4 for L50/ G50, L75/G25 and L100, respectively, after 24 days. The average release rate of BSA from L50/G50, L75/G2 5 and L100 is 3.2, 2.4 and 2.1 wt%, respectively, during the first 24 days.

It has been reported that the release kinetic of BSA grafted to the PLG copolymer follows the degradation rate of the copolymer [19]. Additionally, it has been previously shown that the degradation kinetic of LG based micelles depends on the proximity of water molecules to L and G ester groups [20,21], which is dependent on hydrophobicity of the degradable units [19]. Therefore, the fraction of less hydrophobic G in LG segments has a profound effect on LG unit degradation and consequently the release rate of protein. As a result, by increasing the fraction of G, which is less hydrophobic than L, from 0 to 50%, the the average release rate of BSA increased from 2.1 wt% to 3.2 wt%.

Conclusion

It was shown that by surface modification of silica nanoparticles with short segments of lactide and glylcolide, the controlled release of growth factors can be achieved. Based on the release profile of BSA from surface modified silica nanopartilces, by increasing the ratio of glycolide segment from 0% to 50% with respect to lactide segment, the average release rate of BSA will increase from 2.1 wt% to 3.2 wt% per day. Some of the potential challenges of using nanoparticulate systems for drug delivery applications are the stability of the nanoparticles in aqueous environment, bioactivity of growth factors and controlling release mechanism of growth factors from nanoparticles. Therefore, further studies need to be done on the properties of surface modified nanoparticles as well as controlled release of target growth factors like of BMP-7, TGF-^1 and IGF-Iand their effect on chondrogenic differentiation of MSCs.

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