I've been experimenting since Feb of 2021. It hasn't been easy. I am NOT a typical responder by any metric. I was born super premature with brain damage due to oxygen deprivation and am considered to be somewhat on the spectrum. Later in life after my first cancer I had one of those drug response tests done and it says I am a super rapid metabolizer.
I have a long documented history of things not working right. I did Ketamine therapy for depression and I was taking 650 MG via IV and I was up and walking 30 minutes later. That's not supposed to happen. I can take a half dozen Ambien with zero response. I don't really drink as I don't care for the taste and it's frankly a dedicated job if I want to feel anything from it. To get me drunk would require an amount that would hospitalize some people. Sleeping pills don't work. 50 mg+ injection of Benadryl, no response. I have no response to Opioids..which a great discovery when you're on the oncology floor. I've had two surgical procedures with only local numbing because I cannot be twilighted. If it doesn't require legit going under..then I don't get pain management. I confound the hospital. I had 3 rounds of immensely harsh chemo. I didn't feel a thing, no pain, no discomfort. It was like sparkling water.
I'm telling you this so you can get an idea of just how fucked up I am in terms of drug response.
My medicine of choice is shrooms. But I need a lot. about 4 to 5 grams with rue ie Psilouacha, which to my understanding at least doubles the potency if not more. So I'm probably around 10-12 grams per journey.
I would change myself in a second if I could, but I can't. I just have to deal with things as the immense drug fuck up that I am. I don't get any sacred geometry. That magic hyperspace realm a lot of you go to..I don't have the faintest idea what that is like. Things look a bit brighter..and after a extensive supplementation regimen I can see trees do some odd shit but that's as far as it goes for me in terms of visuals. Internally? Black hole
I've had an immensely traumatic childhood/life. I've been in therapy since grade school and I'm 41, been on meds since the same time roughly. Been on pretty much every cocktail out there with minimal effect. Lithium has been the med of choice for a while but I've been off it for a long time. I used to disassociate heavily as kid, have diagnosed ptsd and whatever else. My oncology therapist says I'm the most fucked person she's ever had. And when the floor staff heard some of my story when I was inpatient, they sobbed. I'm a fucking mess.
I'm pretty much emotionally numb inside, I don't really feel things. I can recall a handful of emotional experiences in my life, that's it. For instance, I know my parents love me, but I don't feel anything, I can't...really connect with people, my mother says I see them like trees. Like objects. The first time I ever felt what seems like love was my first aya trip in May of 2021.
When I do a journey..I can experience..love and joy and grief and..wonder. Emotions and feelings that I can't otherwise access in sober life. It's like I get to cosplay as a human for a day.
I have a progressive chronic permanent illness that cannot be cured and it will eventually result in my death. There's a lot of complications with it. I can't really interact with society. Heavily immune compromised, I've had to live with my parents for the past 5 years in their spare room. Can't be overheated, can't be exposed to sun for more than a few minutes a day. Can't tolerate cold or mold friendly environments. I have damaged lungs, can't do anything physical to any real degree. Tired all of the time.
Psychedelics..are like my window/door to something else outside of my little bedroom. And when I do them I feel great..like perfect, I feel good and energetic...like before cancer type good. It even seems to be effective in rolling back some of the dysfunction/damage that I have. I had massive light sensitivity and a trip last year eliminated that. I don't have to wear sunglasses 24 hours a day anymore. It's far more effective than anything the hospital throws my way.
I started around every 3.5 to 4 weeks. It's been pretty great overall. Most challenging experience was I had the great timing to dose about 2 hours before a mass shooting. That was..not a great time.
Last summer they stopped working as I had advanced thyroid cancer and apparently the thyroid is important to psychedelic trips. Things looked prettier but emotional connection was pretty much gone. Had to get it removed and radiation etc, blah. Post surgery, amazing, it's like a filter was removed. Hospital says I've had it ongoing for years, so ever since I started. So from Aug of 2021 to Oct it didn't really work at. Post surgery, better than it had ever been.
In Jan of this year, I had radiation scheduled and I managed to catch Covid for the first time. I'm considered unvaccinated as my disease requires immune suppression to treat which makes vaccines useless. I tried a trip a week post Covid as my radiation treatment was soon had a really shitty prep and fuck all. Absolutely nothing. But I've read that Covid can really screw with it. So I wasn't surprised by that.
This past Feb was one of the most amazing trips of my life. I had recently lost my aya Shaman Steve Hupp and on that trip I connected with him somehow and it was the most incredible experience.
But ever since then it hasn't really worked. Now..the only change that I can say happened around that time was I vacuum sealed my supply because I was worried about degradation. I cannot think of a single rational reason why that would fuck everything up. I cannot think of any logical reason why that would impact things. The strain I've been using is PE6. My go for a while was Z strain, but I dumped that entire supply into making blue honey which turned out to be a massive mistake. I have old PE6 and GT on hand. I am planning on doing an APE grow..but there's a lot going on atm with my fathers health. We just ruled bone cancer for him, but he's going to need major spine surgery with a decent odds of paralysis. That shit has to be dealt with first before I embark on a new grow.
Sometimes you can just get weak shit..but this has gone on long enough that I think something else is going on.
Lately..it like doesn't kick off. Like a mild afterglow is all I get, and that's after hitting a vape to bring it up. My typical method is lemon tek, though I am planning to experiment with a chocolate bar as I hate drinking that shit.
Maybe something in my health has turned and it's impacting this. If that's the case I'm probably just screwed.
I did a experience a couple days about, 7 grams and change of PE6 with lemon tek. Extremely puffy powder, it just absorbed an entire lemon..it turned to the consistency of oatmeal almost or grits...it was..odd. That was a chore getting down.
It felt like it took forever to go anywhere, normally 40 minutes and you kind of get shoved off the cliff. I had to hit the vape pretty decent to get anywhere.
Normally I have really excellent recall with this stuff. I'm very present. I went to the bathroom to look out the window. It was really hot that day and I wasn't allowed to sit outside, so I went to the bathroom to sit and look out the window. Fucked as it sounds it's the only way sometimes for me to look at nature. I remember looking out the window and then it was dark. I lost time like hours of it. Which has not happened before. The afterglow for me usually lasts for some days..but it was gone that night. Now my mother was explaining just how potentially fucked my father is..so that may have had something to do with it.
I just want my normal emotionally fulfilling experience..

WARNING: SPOILERS GALORE IN HERE, IF YOU HAVEN'T PLAYED ALL THESE GAMES AND DON'T WANT TO BE SPOILED, DO NOT READ!
I've always liked KH, since I first saw Cloud in a KH1 commercial as a kid (I was obsessed with FF7 as a small child), I knew I had to play it.
So I played KH1 religiously in elementary school, heard about CoM but didn't have a gameboy advance so I ignored it, then played KH2, barely understood what was going on, but enjoyed it thoroughly. Continued to ignore the 'spinoffs' because my family was poor and there was no way I could afford all the different portable consoles they were putting these damn games on. Then, finally, bought and played KH3 on release, with my wife.
fast forward to a couple months ago, and after a few debates with my coworkers about the 'quality' of KH3 (I kinda hated it), I decided to sit down and play through the entire series properly, because it was VERY apparent I was missing too much context, and thankfully, 1.5+2.5 final mix, and 2.8 are things that exist now, and are free on PSPlus, so why not? Besides, my wife's ONLY exposure to the series was KH3, so I owed her some context.
Kingdom Hearts 1: What can I say? It's KH1, it's Simple, it's Clean, it's a good fuckin' game when nothing like it really existed at the time. I played on Normal because my wife was watching along, and I didn't want the plot progression to get hung up on me dying to the same boss 25 times. In hindsight, maybe I should've picked a harder difficulty, because I remember spending hours on Maleficent's dragon form as a kid. Hiding out in the bramble where none of her attacks reached me, just chipping away at her HP, but now, I just kinda walked right through her. My biggest complaint with KH1 was against bosses like Ansem's second phase (when you're fighting him 1v1), that KH1's lack of gap-closing options becomes apparent. Bosses like Ansem will frequently blink to the opposite side of the arena, and it takes Sora so long to close the distance, that they've already moved into an attack with i-frames before you can reach them, leading to a lot of missed opportunities for punishes, in my opinion. It didn't help that I accidentally put myself in a sort of 'hard mode' by choosing the Dream Sword, and discarding the Dream Shield, making Sora a glass cannon. But on the flip side, in comparison to later entries the franchise, I really like KH1's more grounded movement, but I'll get more into that later.
I beat the game pretty quickly, so I farmed for Ultima Weapon, as I'd never gotten it as a kid, and then kinda... walked right through Sephiroth, Hooded Figure, Phantom, and Kurt Ziza with little to no effort. Hell, I beat Seph first try, so that was a bit disappointing but I think I did it to myself by grinding out Ultima Weapon first.
Chain of Memories: This was it, my first proper "spinoff" title and I was very excited to get into it, and discover new pieces of the story I missed, and it did NOT disappoint! being properly introduced to the Organization members prior to KH2, finally understanding why Namine is a thing, memories, heart's memories, etc. All to culminate in a proper setup for KH2's opening and to explain why Sora was in a giant flower, etc.
Gameplay however... I knew it was card based because I remember my friend as a kid briefly showing me the game, so that was no surprise, and I honestly did enjoy it enough, the card-based combat was fleshed out enough to give players plenty of room to develop strategies and that's really all I needed from it. What I didn't like, however, was the gameplay loop. It simply got repetitive, and it was obvious that this game was a reskin of a GBA game, hardware limitations included. Every cutscene at the beginning and end of a floor was great, especially towards the end when the climax starts to really ramp up, but getting to those moments was such a slog, so by halfway through the game, I was over it and just wanted the game to end.
Marluxia... omfg. Okay so I played this game on normal, but I guess I screwed up and spent too many levels on max HP, and not enough on deck size, this fight took me for-fucking-ever to beat, I genuinely hated this game by the end, and to top it all off, I misread the endgame prompt and didn't create a "completed save", so I couldn't start Riku's chapter, unless I went back and beat Marluxia again...
358 Days/2: Moving on, even though I own a DS, I'm not buying a copy of this, so I watched the included movie in 1.5+2.5, and honestly, it was great. It added SO MUCH more context to KH2, and it was fun to try and piece together at what moments during KH1 or CoM that certain things were happening in Days. Axel's outright bizarre motives in KH2 finally make sense, Roxas's memories, etc. I will say, though, that Xion confuses me, I really don't understand how Vexen, who constantly talked about his replicas being incomplete, imperfect, etc, was able to make something so powerful, and unique, as Xion. Total outlier of his skills and it never feels properly explained but oh well.
Gameplay's a little light, though.
Kingdom Hearts 2: I was REALLY excited to get back into this one, and it didn't disappoint. With the proper context from CoM and Days, the beginning FINALLY made sense and by the end of Twilight Town, my wife said "Roxas really got done dirty, and I'm mad." To which I agree. Going through this entire game with the intended amount of context was like playing a whole new game, it made a game I already consider to be great, even better. What else can I really say?
When I got control of my character the first thing I remember saying was "jesus christ was the camera always this far away?" I guess I never really noticed it, but with combat being so much more aerial focused, it makes sense. I had never played the Final Mix version of this game, so the Limit Form was a welcome sight, even if it is hilariously broken. Overall, watching Sora's moveset expand as his skills with the Keyblade do, is neat, and reaction commands are a fun mechanic to play with, personally.
However, I did not like how 'floaty' Sora is, and it's a theme that continues through the rest of the series. I died a lot due to getting knocked into the air by a hit, and then just never getting the chance to touch the ground again because Sora takes 10,000 years to float back down to the ground. With no aerial blocking, and air dodging having a longer cooldown then dodge roll, it led to a lot of frustration. Also, special shout-out to "YoU ClEvEr LiTtLe SnEaK!", honestly, fuck Xigbar, I don't know why, but his fight gave me more trouble than any other boss in the entire game. I struggled so much to find an opening to punish him and spent 90% of the fight swinging at the air then getting hit in the back because of how much he blinked around.
I never got around to getting ultima weapon, or beating any secret bosses in KH2, although I did visit Lingering Will and Sephiroth just to see what they were like. I remember I beat Seph as a kid at lvl 65 specifically because of MP Berserk, so maybe I'll go back and try it sometime.
Birth By Sleep: I heard so much about this game as a kid, and even recently, and I knew during my first playthrough of KH3 that Terra, Aqua and Ventus were from BBS, and that they were pretty important characters, but I could've never guessed just how important until I finally played this game. The story for this game was simply great, I started off thinking Terra was the most bland, uninteresting character I'd ever seen (his VO's flat delivery of dialogue didn't help), but by the end, he was my favourite, and honestly, Aqua felt like the third wheel to me, by the end. Like sure, after completing the 4th 5th and 6th episodes, I felt bad for her, as she was clearing going through A LOT, but while Terra and Ventus both had VERY clear stakes in the story, Aqua just felt kind of... there. Her ties to the plot weren't nearly as strong and her story was more relegated to "watching her 2 friends get fucked by destiny and plot". However, Aqua at the end being responsible for Castle Oblivion, along with the clarification on what Org. 13 was looking for in CoM, was such an awesome reveal and my wife and I were losing our collective shit over it.
Oh, and of course, finding out what exactly the Lingering Will was, was pretty damn cool, too!
Fun fact, this is the first game I played on Critical, everything was on Normal until this point. I heard it was one of the easier games to play on Critical, so I figured why not. By the time I finished Terra's arc, I was begging to go back to CoM to fight Marluxia. I cannot express enough just how much I DESPISE this game, in a gameplay sense. Ignoring the obvious things like playing through the same worlds 3 different times and how tiresome that gets, the combat was so blood-boilingly infuriating, I wondered if I was going to actually finish the game. The command system, I hate it. "What? You wanted to cast Cure because you're about to die? Well, too bad, the enemy with 0 warm up frames in his attack just hit you at the same time you pressed X, so your Cure goes on cooldown now, fuck you." That's not even getting into the terrible end-lag in your own attacks, and just how sluggish everything feels. Critical mode aside, this game even on Normal would've pissed me off for all the same reasons, I was so relieved to be done with it.
Re:Coded:>! I was told this one wasn't necessary, and almost skipped it, but decided against it and I'm kinda glad I did. I only watched the movie, but I hear the gameplay itself is pretty solid, so maybe one day I'll check it out. The plot is self-contained, and interesting to watch, and it does get referenced a few times in KH3, so it didn't really feel like a waste.!<
Dream Drop Distance: I was genuinely lost for a while, plotwise, in this game. I felt like I'd missed an entry somewhere that'd apply context, but no, Yen Sid is just not great at explaining things. By about halfway through the game I was caught up and understood what was going on, but the first hour or so I was just along for the ride, hoping I'd understand it later. I enjoyed what DDD was doing, and it felt like, had it been a console release, this could've easily been KH3 with just how much direct impact it has on the plot compared to other entries. Honestly, watching Riku slowly, but surely, get his act together throughout the series has been a treat, and it all comes to a head in this game, he finally moves on from rival/edgebaby to a proper equal to Sora, so much so that if they were to have a gentleman's duel just to test their strength, I couldn't confidently say who would win.
The worlds were hit or miss, seeing Notre Dame was cool, but honestly who the hell asked for Tron: Legacy? I will note that, especially for worlds like Tron, I didn't really feel like I was part of the plot like in KH1 and 2, but more like I was just watching the plot progress from the sidelines. It was jarring.
But the biggest disappointment by FAR was Fantasia. Before I even booted up this game, I was talking about how cool it would be if KH ever visited the world of Fantasia just due to how unique of an experience it would be. I imagined something along the lines of a rhythm game like Crypt of the Necrodancer or Metal Hellsinger, just a neat gimmick tied to that world's music that kept you in line with the beat of the song. So, when I watched the opening to DDD and saw the very clear reference to Fantasia, I got excited, only to be completely let down by it being just another world, except your hits play random instrument noises, and essentially a barebones prototype to Melody of Memory as a reaction command. Honestly, this world was completely wasted on a portable KH entry, it should've been saved for a console entry where it could've been realized more fully, rather than confined to the limitations of a portable console. Oh well, maybe in KH4.
I also played this game on Critical and hooboy did I feel it, but first, the basics. Flowmotion sucks. Thanks for coming to my TED talk. Seriously, I get what they were going for, but I honestly hated how disruptive flowmotion ended up being. Air dodge out of the way of something, well, you touched a wall, so now you gotta brace against it and get ready to bounce off. It's not that it was ruining my gameplay or anything, it was just annoying because it'd remind me of its existence at the most inconvenient times, and it was broken, like, hilariously broken. I definitely abused flowmotion to get through some encounters early on, when I didn't have many tools at my disposal yet, and even in the World that Never Was, I managed to use flowmotion to get ontop of, and over a building to get to the other side, which triggered a cutscene I don't think I was supposed to see yet, because suddenly I was talking to Roxas with NO context, so there's a good chance I skipped a scene or two by doing this purely because the game let me, and I wanted to see what was at the top, if anything (I originally thought I was gonna find some way out of bounds or something).
The Dreameaters genuinely don't bother me. I actually kind of enjoyed levelling them up as a means to acquire abilities and passives. The only thing that unironically kept me from grinding their stats more is that irritating fucking song that plays whenever you play minigames with them. I despise it. With all my heart.
The drop-out mechanic was annoying as hell in some instances, especially when, towards the end of the game, I did a TON of farming on Sora for Droplets, to give Riku a ton of bonuses to help with progression, only for the game to immediately FORCE me to drop from Riku, back to Sora, just as soon as I got control of him, causing me to hard reset the game, reload my save, and actively backtrack as Riku to get an actual use out of the bonuses I saved up for him.
The combat was, well, I wasn't enthused to see the Command List back, and I suffered all the same frustrations that BBS gave me, with only MARGINAL improvements in gameplay. The only saving grace was enemies actually had warm-up animations on attacks, giving you a brief head's up instead of just randomly bursting into a combo with no warning.
Anti-Nightmare Sora was the first fight that I genuinely gave up on. even Xigbar in KH2, Terranort in BBS, Terranort again in BBS, Vanitas, all of them I managed to push through. But this fucker? No, I had to walk away for a day, come back, grind my eyes out, and when I FINALLY beat him, the game then beat me over the head with Young Xehanort...
...Then I discovered Balloonra and slept through the rest of game.
Fragmentary Passage: Not much to say other than it's a tech demo for KH3, plot wise, it was interesting to see where Aqua's story lines up with the end of KH1, and where she's at both literally, and mentally, by KH3.
X: Back Cover: Whoever wrote the dialogue for Master of Master's needs a raise and a personal statue outside SE HQ because this guy is hilarious! His deliveries are genuine, quick, witty, and not at all the stilted, oddly paced dialogue I've come to expect from the KH series, and well, everything Square does outside of FF14 and 16. I would not be surprised to find out his dialogue was written in English first, rather than Japanese.
Anyway, I'm really mad I have to either play, or watch someone else play, a gatcha game to get the rest of the context here, because this movie was fascinating, and these characters all seem to be pretty damn important to KH4, so it's a shame.
Kingdom Hearts 3: Back (almost) full circle! Just like with KH2, coming into KH3 was full context was such a breath of fresh air! Everything made more sense, and I had a clear understanding of the character's motives and goals throughout the story. Again, what's there to say, really?
Well... the worlds kinda suck. Olympus being relegated to a tutorial felt like a slap in the face, and overall, the entire world felt bland. It looked like a generic Unreal Engine open world environment, with Hercules pasted in it. Nothing about the open wilderness outside the city really looked like it belonged there, it didn't fit Hercules' art style in the slightest. Compared to, say, Kingdom of Corona, where everything felt more visually on brand, granted, Tangled is a CGI movie, Hercules isn't, I understand that the translation is harder to make but it really didn't look like much effort was taken at all for 2/3rds of Olympus.
Arendelle was "Fall of a mountain simulator" and I was absolutely sick of it. It also epitomized my problems with DDD's worlds in that, when you really think about it, you do ABSOLUTELY NOTHING in Frozen's plot. You are sidelined the entire time, with Sora and the gang essentially climbing back up a mountain, being told "hey here's what you missed, okay bye!" Before being yeeted back off again, to rinse and repeat ad nauseum until you fight a boss that just doesn't feel earned, which is a damn shame because Skoll is one of the coolest looking fights in the game. There's no participation, the plot simply ignores you the whole time and does it's own thing until you leave.
Pirates and Big Hero 6 felt hilariously short. If you have no intention to explore the open world (there is nothing worth exploring in Sanfransokyo, so this doesn't help) The worlds feel like you're in and out in 20 minutes.
Shoutout to Monsters Inc and Toy Story, you definitely felt like you were a part of the world, and a part of the story happening within it.
The end of this game was phenomenal, seeing everything you've been watching build up, finally come all together, the reunions, the realizations, the callbacks, all of it was such a MASSIVE payoff that was completely lost on me the first time, when the only games I had played were KH1 and KH2, and not since they were new.
So, as with every other game after BBS, I played this one on critical, as I was told it was simply a more rewarding experience, and I honestly agree. Sure, I had a lot of frustrations (Sora's floatiness being paramount) but you can finally BLOCK IN MIDAIR, and BLOCK OR DODGE MID COMBO! If KH2 had these mechanics, it'd be hands down the best game in the whole series, period. I wish they'd thought of it sooner, almost every entry in the series would have benefited from this.
I did spend my fair share of time absolutely raging at the final boss gauntlet, however, and really, my biggest issue was just visual clarity (if you remember Xehanort's meteor attack, then you probably know). But I beat it and was very satisfied with how Xehanort's arc turned out. KH3 is a great game.
Re:Mind: I got this DLC half off, and even still, I'm a little mad I spent money to spend most of my time rewatching the cutscenes I JUST WATCHED from the maingame. The game reached a point where it started truncating the cutscenes to get to the new info quicker and I couldn't help but wonder why the hell they didn't do that to begin with... but ultimately, the DLC was good, a glorified boss gauntlet, sure, but good. Explaining Sora's disappearance was welcome and started setting up context for KH4.
The data fights... as of writing this, I have completed 11/13 of the fights. I'm missing Xion, Xehanort, and of course, Yozora. Only reason I haven't done more is a sudden lack of free time to do so. To reiterate, I'm on Critical, and these fights, while some have absolutely pissed me off to no end (lookin' at you, Vanitas), these fights have solidified KH3 as probably my favourite game in the franchise, purely on gameplay alone, and one of my favourites of all time. The combat FEELS rewarding, (usually) failure doesn't feel like you were cheated by some BS mechanic, but feels like something you know YOU did wrong, and need to correct for the next attempt. I will be going back to do the rest of the fights, as terrifying as they look, because so far, it's been genuinely fun to figure out the puzzles behind these fights and put my skills to the test.
Melody of Memory: Kingdom Hearts rhythm game! Neat!
Well, kinda. It wasn't a very satisfying game to play in comparison to something like Guitar Hero, but it was enjoyable to listen to my favourite KH songs (shout out to The Encounter! Don't care how bullshit the boss is, if it's using that song, I can't even get mad, it's just too much of a bop!) But I did not appreciate yet another rehash of the plot as the main gameplay incentive.
Thankfully that game is pretty damn short, so getting to the end, where the actual meat and potatoes is, in terms of new plot, was pretty painless and overall, worth it. More context for KH4, Kairi deciding to train under Aqua(!), and the somewhat harrowing implications that Xehanort knew about the 'unreality'.
And that's it. I guess I'm done with KH until 4 releases, or that AR mobile game. I've been typing for a very long time now and I'm not sure how to close this out other than... uh, thank for reading, can't wait to hear all about how wrong or right I am about your favourite entries!

Hey All,
I am a PGY-3 neurosurgery resident at Northwell, and we are currently running a crowdfunding campaign to raise money to create a low-cost biomechanics testing apparatus for Northwell. If successful, this campaign will provide the funds to begin construction of the skeleton of our testing apparatus. There are no options for biomechanics testing near our facilities, and commercially available equipment costs north of $200K, compared to our design which would cost approximately $40K. Any bit helps. Happy to answer any questions about the project in the comments.
I cant include the link, but the crowdfunding campaign can be found by going the the "Experiment" website and searching biomechanics. Its the only one that shows up.
Thanks for reading!

Hey All,
I am a PGY-3 neurosurgery resident at Northwell, and we are currently running a crowdfunding campaign to raise money to create a low-cost biomechanics testing apparatus for Northwell. If successful, this campaign will provide the funds to begin construction of the skeleton of our testing apparatus. There are no options for biomechanics testing near our facilities, and commercially available equipment costs north of $200K, compared to our design which would cost approximately $40K. Obviously we are all living on the cheap, but any bit helps. Happy to answer any questions about the project in the comments.
I cant include the link due to the subreddit auto-moderating my post, but the crowdfunding campaign can be found by going the the "Experiment" website and searching biomechanics. Its the only one that shows up.
Thanks for reading!

hi fellow lab rats,
thanks in advance for your help with this. i feel like i must be missing something obvious, so an outside perspective to my own will be very useful.
TLDR; my protein library expressed great in E. coli colonies on agar plates, but when i cultured these colonies to prep and test the protein variants, expression was so bad i got functionally no data. what could cause this?
full explanation:
i recently made a library of a red fluorescent protein (RFP), chemically transformed that library into T7 Express (a commercial line of highly chemically competent E. coli), and plated my transformants on agar plates with appropriate antibiotics. i grew these plates at room temp (just out on my lab bench) over this past weekend. on monday, the plates looked fabulous -- i could easily tell that my protein was expressed well because my colonies were very pink in normal lighting and fluoresced brightly under illumination by a ~500nm wavelength light box. given that my plates looked good, i picked single fluorescent colonies into wells of 96-deep-well culture blocks containing antibiotic autoinduction media, one colony (and therefore one protein variant) per well. i also picked a few wells of the library template into these blocks to use as a control. i then cultured these blocks at 30C for ~48hrs and shaking at ~300rpm. today (wednesday) i took them off the shaker to prep and test my protein variants to see if i got any hits, and noticed immediately that the bacteria pellets, including those expressing my control, were barely pink. i prepped the proteins anyway and got unusable data because they expressed so poorly.
i have been evolving this RFP for the past year; this is my tenth library of this RFP, and i use the same workflow described above for every library with few issues until now.
does anyone have any ideas as to why my protein would express well on agar plates but not in culture? i double checked that the autoinduction media i used was in-date, which it is. i also checked that the shaker was actually on before i left on monday, checked it again on tuesday, and it was still shaking this morning, so i don't think it was stopped at any point during the culture period.
my plan for now is to just re-plate the library (i always save the library DNA for this very reason), re-do the experiment, and hope for the best, but any advice or insight will be much appreciated. thank you!!

My 16 yr old niece is about to turn 17 and will be a junior this fall. She is the oldest of 3. Her parents have always treated her like a slave while the other 2 get away with murder. Her mom is a narcissist, since the day i met her. Her mom has always been jealous of the attention my niece gets from the family bc she is sweet and has a great personality unlike her mother. She has even threatened her in front of everyone for no reason. Recently she got a bf and a job. Her parents have found out her and her younger brother have been smoking and went ape on her but not her brother and they drug test her every week but not her brother. Anyways they told her today her relationship with her bf isn't healthy bc they talk all the time on the phone when they are not together and they are revoking her cell phone privileges. Now her younger brother has had a gf off and on and they never stalked him like they do her. I have offered her a phone on my phone plan and room if she ever wants to move out and access to my car whenever she needs it. The reason for that is because they have not set her up for success, birth control, college prep, nothing. I do not want her to end up like me pregnant senior year and dropping out of college, which is what her mom secretly wants for her. AITA for going behind their back and offering these things to my niece?

Background: I work in a very somewhat niche and technical role. I have an engineering background with about 10 years worth of experience. Most of my prior roles have been in R&D/technical roles where I would be in the weeds as they say.
My current role is an advisory/consulting role, I started here 10 months ago and have been promoted once already. I am the only one with a technical background - all my colleagues are from business/project management backgrounds. Without sounding big-headed and arrogant, the work is easy, there is no real engineering here and in a typical week I probably do about 4/5 hours of real work. Additionally, the commercial guys are struggling to sell the services of the firm, especially within the UK (I think they may have overestimated the UK demand). Basically, I am bored and spend most days watching videos online, doing personal hobbies or just taking extra long lunches.
Recently, I got headhunted by another company, although the role is still advisory/consulting, on the face of it the role is a lot more involved and has a good balance between the concepting, design phases and the physical manufacturing/testing.
Here is the situation: The new company have offered a very generous salary (more than 50% increase on my current salary), with an uplift at the end of the probation and all the usual benefits. The role is fully wfh with client site travel at the start and end of projects. The company is a Chinese company who have locations in the UK, Malaysia, Hungary and China. My role would require me to work quite closely with my Chinese colleagues.
My current employer doesn't want to lose me and has offered to up my salary (although it's still 5k short of the Chinese offer), has looked into a secondment/relocation to one of our other offices and various other incentives.
My questions:

1. Given the current geopolitical issues and the general stereotype of China, would you take the role?
2. What should I look out for, and be worried about in the long term?
3. Should I be running for the hills?
4. Should I just suck it up and have an easy life/easy job?

Thoughts and Opinions are welcome as I am struggling to make a choice.

​

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Table of Contents
What is Tunnel Engineering?
What is the purpose of the Tunnel?
What is Tunnel design?
What is the well-known tunnel design Software?
What are the methods of Tunneling?
Cut and Cover Method
Bored Tunnel Method
Clay Kicking Method
Shaft Method of Tunnel Construction
Pipe Jacking Method
What are the different types of tunnels?
Tunnel for traffic
Tunnel for Hydropower
Tunnel for Public Utility

What is Tunnel Engineering?

Underground construction and tunnel engineering are genuinely interdisciplinary processes, requiring knowledge and skills from civil engineering, geological engineering, and mining engineering, as well as mechanical engineering, geophysics, electrical engineering, geology, etc.
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Students must learn to design, build, rehabilitate, and maintain structures peculiar to the underground space, and the use of tunnels for commercial, transportation, water, wastewater, and utility, etc in addition to typical construction procedures.

What is the purpose of the Tunnel?

Tunnels are passages beneath the ground that are utilized for transportation. They might be used to transport freight and passengers, as well as water and sewage. Beyond a certain depth, tunnels are more cost-effective than open cuttings. During the construction stage, tunnels normally undisturbed the surface activities like live traffic, open recreational activities, etc.
Tunnels are considered to be less expensive than bridges or open steep earthwork cuttings for highway projects or transporting public utility services such as water and wastewater or gas line. The inability of these structures to be built out of natural resources such as rock and soil leads to their failure.
Geological factors are expected to play a significant effect on their long-term stability. Aspects that are extremely important are such as ground and sub-ground conditions, construction sequencing, construction period, construction cost, etc. All these factors play a very important role in determining the feasibility of a tunnel project.
Read More:
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What is the Function of Bridge Foundation or Pier and its Types? 5 Important Points

What is Tunnel design?

Accepting the inherent ambiguity linked with subterranean conditions that can be defined but never really known is a prerequisite for the tunnel design and construction process.
A typical tunnel project starts from the exploration of suitable ground conditions and then testing the engineering parameters, then moves to the selection process of tunnel excavation methods, then moves to the development of liner design parameters, and estimating surface settlements, and finally covers their effects on adjacent structures.
It uses various case studies of successfully executed completed projects to frame out the risks and mitigation strategies, lesson learned activities for the proposed project.

What is the well-known tunnel design Software?

• Tunnel Design Software
• PLAXIS3D tunnel software
• Flac3D

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What are the methods of Tunneling?

Cut and Cover Method

Shallow tunnels are often built using the cut and cover method of tunnel construction. This approach involves excavating a trench in the ground and covering it with support capable of supporting the load. There are two methods for the cut and cover method. One is the bottom-up method, in which a tunnel is excavated beneath the surface and supported by the ground.
Another way is the top-down method, in which the side support walls are built first, either via slurry walling or contiguous bored piling. The roof is then installed on top of the walls, and excavation is completed. Finally, the base slab is built. The majority of underground metro rail stations are built utilizing the cut-and-cover approach.

Bored Tunnel Method

The method of using a bored tunnel is cutting-edge expertise. Tunnel boring machines are used for this method, which works automatically and makes the entire tunnelling procedure easier. It is also a more efficient and effective way of tunnel construction in urban, heavy trafficked locations where traffic diversion, and traffic management during construction is a highly difficult and expensive task.
Tunnel boring machines (TBMs) come in a variety of shapes and sizes to meet various ground conditions. These machines can work in challenging environments, such as below the water table. TBM can work in below-water-table circumstances because of a specific pressurized container.

Clay Kicking Method

This procedure is utilized when there is a lot of clay in the soil. This is a traditional approach that is still used for little jobs such as installing sewage pipes. In this procedure, a hole is dug into the earth, and then a depth tunnel is constructed by the clay kicker, which is mounted on a plank at a 450 angle. Under the clay kicker foot, there is a digging tool.
This is well-known since it was the method employed by Englishmen during World War I to bury mines.
Read More:
How to learn STAAD Pro? 7 Important Points
How do you answer tell me about yourself in an interview? 5 Important Points

Shaft Method of Tunnel Construction

In this method, the tunnel is constructed at a greater depth from the ground surface. The shaft is placed to a depth where the tunnel is required. The shaft is a long-term construction that works well with concrete walls. TBMs are deployed to excavate tunnels to the appropriate depth.
Tunnels have shafts at both ends. If the tunnel is excessively long, intermediate shafts are constructed. These shafts can be utilized for ventilation purposes and for emergency exits once the construction of the tunnel is completed.

Pipe Jacking Method

The pipe jacking method is recommended to build tunnels beneath existing facilities such as roads and railways. Hydraulic jacks are deployed to drive specifically designed pipes into the ground. Tunnels can have a maximum diameter of 3.2 meters.

What are the different types of tunnels?

Tunnels can be grouped into three main categories based on their intended use:

• Tunnels for traffic
• Tunnels for hydropower
• Tunnel for public utilities

Tunnel for traffic

Tunnels are constructed to transfer traffic load from surface to subsurface channels for a brief period of time in order to facilitate congestion-free traffic movements. Traffic tunnels are structures that allow traffic to move at a chosen pace, with optimum ease, and at a minimal cost.
There are three types of traffic tunnels:

• Railway tunnels,
• Highway tunnels, and
• Pedestrian tunnels.

Tunnel for Hydropower

These tunnels are constructed for hydropower generating, as the name suggests. They are hammered into rocks, allowing gravity to convey the water.

Tunnel for Public Utility

These tunnels are constructed for a specific function, such as the disposal of municipal trash, the transportation of pipelines, cables, and oil supplies, as well as water. In today's heavily populated cosmopolitan cities, they are also constructed for underground parking and storage.
Tunnel Engineering online courses Being a unique engineering discipline, Tunnel Engineering courses are expensive and are very few available in the market. Following are some well-known courses in Tunnel Engineering:

• Postgraduate course in Tunnel Engineering
• Certification course in Tunnel Engineering
• NPTEL course on Rock Mechanism

Gcelab.com also offers a unique Tunnel Engineering online course. The course covers various key aspects of Tunnel Engineering and assists the student to learn Tunnel Engineering and move forward in the right direction to make Tunnel engineering an aspiring career option.
I hope this blog offers you adequate information about Tunnel Engineering and Tunnel Design.
Please feel free to like, share and comment about it.
Admin, gcelab.com Please see our Pillar Post to know why we founded gcelab.com.
Read More:

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• How to Become a Road Safety Auditor: 10 Important Points

Allhackgecko.com ---------------------- Digital forensics or digital forensic science is a branch of cybersecurity focused on the recovery and investigation of material found in digital devices and cybercrimes. Digital forensics was originally used as a synonym for computer forensics but has expanded to cover the investigation of all devices that store digital data.
What is Digital Forensics?
As society increases its reliance on computer systems and cloud computing, digital forensics becomes a crucial aspect of law enforcement agencies and businesses. Digital forensics is concerned with the identification, preservation, examination, and analysis of digital evidence, using scientifically accepted and validated processes, to be used in and outside of a court of law.
While its roots stretch back to the personal computing revolution in the late 1970s, digital forensics began to take shape in the 1990s and it wasn't until the early 21st century that countries like the United States began rolling out nationwide policies.
Today, the technical aspect of an investigation is divided into five branches that encompass seizure, forensic imaging, and analysis of digital media.
What is the Purpose of Digital Forensics?
The most common use of digital forensics is to support or refute a hypothesis in a criminal or civil court:

• Criminal cases: Involving the investigation of any unlawful activity by cybercriminals. These cases are usually carried out by law enforcement agencies and digital forensic examiners.
• Civil cases: Involving the protection of rights and property of individuals or contractual disputes between commercial entities were a form of digital forensics called electronic discovery (eDiscovery).

Digital forensics experts are also hired by the private sector as part of cybersecurity and information security teams to identify the cause of data breaches, data leaks, cyber attacks, and other cyber threats.
Digital forensic analysis may also be part of incident response to help recover or identify any sensitive data or personally identifiable information (PII) that was lost or stolen in a cybercrime.
What is Digital Forensics Used For?
Digital forensics is used in both criminal and private investigations.
Traditionally, it is associated with criminal law where evidence is collected to support or negate a hypothesis before the court. Collected evidence may be used as part of intelligence gathering or to locate, identify or halt other crimes. As a result, data gathered may be held to a less strict standard than traditional forensics.
In civil cases, digital forensic teams may help with electronic discovery (eDiscovery). A common example is following unauthorized network intrusion. A forensics examiner will attempt to understand the nature and extent of the attack, as well as try to identify the attacker.
As encryption becomes more widespread, the forensic investigation becomes harder, due to the limited laws compelling individuals to disclose encryption keys.
What is the Digital Forensics Investigation Process?
There are a number of methodologies for the forensic process, which define how forensic examiners should gather, process, analyze, and extract data. Digital forensics investigations commonly consist of four stages:

1. Seizure: Prior to actual examination, the digital media is seized. In criminal cases, this will be performed by law enforcement personnel to preserve the chain of custody.
2. Acquisition: Once the assets are seized, a forensic duplicate of the data is created, using a hard drive duplicator or software imaging tool. Then the original drive is returned to secure storage to prevent tampering. The acquired image is verified with SHA-1 or MD5 hash functions and will be verified again throughout the analysis to verify the evidence is still in its original state.
3. Analysis: After the acquisition of the evidence, files are analyzed to identify evidence to support or contradict a hypothesis. The forensic analyst usually recovers evidence material using a number of methods (and tools), often beginning with the recovery of deleted information. The type of data analyzed varies but will generally include email, chat logs, images, internet history, and documents. The data can be recovered from accessible disk space, deleted space, or the operating system cache.
4. Reporting: Once the investigation is complete, the information is collated into a report that is accessible to non-technical individuals. It may include audit information or other meta-documentation.

What Tools Do Digital Forensic Examiners Use?
In the 1980s, very few digital forensic tools existed, which forced forensic investigators to perform live analysis, using existing sysadmin tools to extract evidence. This carried the risk of modifying data on the disk which led to claims of evidence tampering.
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The need for software to address this problem was first recognized in 1989 at the Federal Law Enforcement Training Center and resulted in the creation of IMDUMP and SafeBack. DIBS, a hardware and software solution, was released commercially in 1991.
These tools create an exact copy of a piece of digital media to work on while leaving the original disk intact for verification. By the end of the 1990s, the demand for digital evidence meant more advanced tools such as EnCase and FTK were developed, allowing analysts to examine copies of media without live forensics.
There is now a trend towards live memory forensics using tools such as WindowsSCOPE and tools for mobile devices.
Today, there are single-purpose open-source tools like Wireshark, a packet sniffer, and HashKeeper, a tool to speed up the examination of database files. As well as commercial platforms with multiple functions and reporting capabilities like Encase or CAINE, an entire Linux distribution designed for forensics programs.
In general, tools can be broken down into the following ten categories:

1. Disk and data capture tools
2. File viewers
3. File analysis tools
4. Registry analysis tools
5. Internet analysis tools
6. Email analysis tools
7. Mobile devices analysis tools
8. Mac OS analysis tools
9. Network forensics tools
10. Database forensics tools

What are the Different Branches of Digital Forensics?
Digital forensics is no longer synonymous with computer forensics. It is increasingly concerned with data from other digital devices such as tablets, smartphones, flash drives, and even cloud computing.
In general, we can break digital forensics into five branches:

1. Computer forensics
2. Mobile device forensics
3. Network forensics
4. Forensic data analysis
5. Database forensics

What is Computer Forensics?
Computer forensics or computer forensic science is a branch of digital forensics concerned with evidence found in computers and digital storage media. The goal of computer forensics is to examine digital data with the aim of identifying, preserving, recovering, analyzing and presenting facts and opinions about the digital information.
It is used in both computer crime and civil proceedings. The discipline has similar techniques and principles to data recovery, with additional guidelines and practices designed to create a legal audit trail with a clear chain of custody.
Evidence from computer forensics investigations is subjected to the same guidelines and practices as other digital evidence.
What is Mobile Device Forensics?
Mobile device forensics is a branch of digital forensics focused on the recovery of digital evidence from mobile devices using forensically sound methods.
While the phrase mobile device generally refers to mobile phones, it can relate to any device that has internal memory and communication ability including PDA devices, GPS devices, and tablets.
While the use of mobile phones in crime has been widely recognized for years, the forensic study of mobile phones is a new field, beginning in the late 1990s.
The growing need for mobile device forensics is driven by:

• Use of mobile phones to store and transmit personal and corporate information
• Use of mobile phones in online transactions

That said, mobile device forensics is particularly challenging due to:

• Evidential and technical challenges such as cell site analysis which makes it possible to determine roughly the cell site zone from which a call was made or received but not a specific location such as an address
• Changes in mobile phone form factors, operating systems, data storage, services, peripherals, and even pin connectors and cables
• Storage capacity growth
• Their proprietary nature
• Hibernation behavior where processes are suspended when the device is off or idle

As a result of these challenges, many tools exist to extract evidence from mobile devices. But no one tool or method can acquire all evidence from all devices. This has forced forensic examiners, especially those who wish to be expert witnesses, to undergo extensive training to understand how each tool and method acquires evidence, how it maintains forensic soundness, and how it meets legal requirements.
What is Network Forensics?
Network forensics is a branch of digital forensics focused on monitoring and analyzing computer network traffic for information gathering, legal evidence, or intrusion detection.
Unlike other branches of digital forensics, network data is volatile and dynamic. Once transmitted, it is gone so network forensics is often a proactive investigation.
Network forensics has two general uses:

1. Monitoring a network for anomalous traffic and identifying intrusions.
2. Law enforcement may analyze capture network traffic as part of criminal investigations.

What is Forensic Data Analysis?
Forensic data analysis (FDA) is a branch of digital forensics that examines structured data in regard to incidents of financial crime. The aim is to discover and analyze patterns of fraudulent activities. Structured data is data from application systems or their databases.
This can be contrasted to unstructured data that is taken from communication, office applications, and mobile devices. Unstructured data has no overarching structure and analysis, therefore, means applying keywords or mapping patterns. Analysis of unstructured data is usually done by computer forensics or mobile device forensics experts.
What is Database Forensics?
Database forensics is a branch of digital forensics related to databases and their related metadata. The cached information may also exist in a server's RAM requiring live analysis techniques.
A forensic examination of a database may relate to timestamps that apply to the update time of a row in a relational database that is being inspected and tested for validity to verify the actions of a database user. Alternatively, it may focus on identifying transactions within a database or application that indicate evidence of wrongdoing, such as fraud.
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https://www.khanacademy.org/test-prep/mcat/biomolecules/amino-acids-and-proteins1/v/special-cases-histidine-proline-glycine-cysteine
Isn't that true for other amino acids with a charged side chain too? Like leucine? Its carboxyl group is deprotonated and amine group is protonated and side chain is protonated at physiological pH?

I take my second attempt on Friday for the Sec+ exam and have been studying my ass off the last couple of days (also ADHD is a big ol hoe for studying).
I have been watching Messer at 1.5x, reviewing my ports, watching some Cyberkraft PBQ and mostly using the Comptia Sec+ Test Prep App for testing.
I ask what else is there that I can study? Im still very nervous. Im really not trying to fail this test again and get my life moving again..

​
https://preview.redd.it/v4slzezucn4b1.png?width=750&format=png&auto=webp&s=b0bb46dcd5962119d0df7a5856ffabf64500051e
The Day Before Your Exam: Avoid Cramming
On the day before your GRE, it is very tempting to try to cram in as much studying as possible. Resist this urge! Chances are, you’re not going to learn a bunch of new GRE material in that final day. Furthermore, you don’t want to stress yourself out or burn the midnight oil right before your exam.
Instead of cramming, do some mixed practice (not hundreds of questions), review some relevant study notes (not all of them), and quiz yourself with your vocab flashcards (not hundreds of them). Keep your studying relatively light, rather than make the entire day about the GRE.
In fact, the day before your exam is the perfect time to do something fun and relaxing. Go for a long bike ride or grab dinner with a friend. Make sure you’ve got everything in order for test day, and get a good night’s sleep. If, in the hours leading up to your exam, you’re scrambling to master new topics or do every last practice question you can find, you’re probably not going to head into test day in the “calm but ready” state that makes for peak performance.
Reach out to me with any questions.
Warmest regards,
Scott

​
https://preview.redd.it/ej5dx88dcn4b1.png?width=750&format=png&auto=webp&s=ad1ab74a6c1014e43e4785dc28aea777c38c7908
The Day Before Your Exam: Avoid Cramming
On the day before your GRE, it is very tempting to try to cram in as much studying as possible. Resist this urge! Chances are, you’re not going to learn a bunch of new GRE material in that final day. Furthermore, you don’t want to stress yourself out or burn the midnight oil right before your exam.
Instead of cramming, do some mixed practice (not hundreds of questions), review some relevant study notes (not all of them), and quiz yourself with your vocab flashcards (not hundreds of them). Keep your studying relatively light, rather than make the entire day about the GRE.
In fact, the day before your exam is the perfect time to do something fun and relaxing. Go for a long bike ride or grab dinner with a friend. Make sure you’ve got everything in order for test day, and get a good night’s sleep. If, in the hours leading up to your exam, you’re scrambling to master new topics or do every last practice question you can find, you’re probably not going to head into test day in the “calm but ready” state that makes for peak performance.
Reach out to me with any questions.
Warmest regards,
Scott

Newby to saltwater.. sorry you probably see that a lot
Magnesium with Salifert NEW test kit. 2 steps not 3. 2ml of test water to vial Add 5 drops water Drops are bluewater turns blue Swirl for 5 seconds, water is still blue
Get dropper 1ml dropper fill to 1ml with Mg 2 solution Drop 1 drop at a time, instruction say until blue or gray what ever comes first. The blue never changes. I repeated first try was ROD water i am prepping for the tank Then bottled water. Then tap, then water from my freshwater tank. Same results blue as soon as I add 5 drops of step 1. Adding step 2 changes nothing. So does this mean test is broke or very source is Magnesium off the chart. If the testing is bad, I see no way to contact salifert. Guess I return to seller?
OR Does magnesium not matter?

I would like to say thank you beforehand, and just be honest.
I came to the US four years ago, didn't know how to speak English. We had political persecution in our homeland (an Hispanic country), so it was either flee or die. We got the asylee status pretty quick, but we haven't get our permanent resident card yet.
I go to a STEM magnet public school in Florida. We have less than 500 hundred students. Stats: 3.9UW/4.5W (might increase over the summer based on some test scores and my over summer AP). SAT:1350 (I know is low for some colleges, but I got into a foundation that can provide test practice over the summer, and I also re took it in June 3rd so that also might increase it as well). Class:7/101 (again, might increase). So far 10 AP (incl AB and C: Mechanics) and three DE classes with UF (I'm also planning to take Calc 2 and comp sci with c with them for the first semester) Only 2 non-honors courses in my HS transcript, the rest are Honors, AP, or DE
Awards: AP schoolar with Honors (might get with distinction over the summer) Academic letter Questbridge college prep scholar Pin Award (an school award) I might have other that I don't remember Sunrise scholar
Experience: -I did a internship with a partnership in my state. In this internship, with peers, we created a detailed business plan of like 15 pages long describing an entrepreneur business idea and all the implications from market implications to costs. -Held jobs during my last two summer as well. ADT and Adidas. -Intern at a company (would say name but don't want to get doxx) that simulates the lunar and Martian soil. Is a really interesting idea and multiple research papers are being published using their simulants. Companies like NASA and other big space companies use them. -Intern in a quantum computing research internship through a program made by graduates students at MIT, Berkeley, and others. I got in by having good grades and a final project for my year long quantum computing course with them. The internship is with a UF professor and their graduates students. -Participated in one proposal for an experiment for (huge space company) that would be send to orbit. Although my group didn't get it. Our school ended up having two experiments. -Participated in the NASA tech rise contest but also didn't get it. - Created a tutoring club in my school that pairs students with other students. The tutors get community hours. - chairman of finances for my interact club -NHS (my club started as a NHS committee) - attorney in my local teen court program - teenager ambassador in my library system - college board embassador.
Intended major: physics and nuclear engineering
Dream school: Caltech Would be happy to get into any big T20 with a good physics program (that leads towards graduate school, since I want to be a professor)
Safeties: some schools in my home state
I would be grateful if I can get any input on this. Thank you
Edit: forgot to mention low income, reason I have questbridge and sunrise. I will also apply considering if they have full based need aid