#and triton for an even less split second

[grape-jucie-dog]

Someone needs to stop me from trying to host a M.A.P nobody will join. 💀

[attina-the-responsible]

How to Tell Your Family You’re Moving Across the World According to Mermaids ~*~ [Merfam]

In which Attina and Panic make a big announcement...[takes place at the beginning of the summer.]

@arista-the-musical​, @aquata-the-champ​, @alana-the-brilliant​, @melody-the-unwritten​, @panicked-nights​

[tw -- none]

ATTINA: Attina had cooked a nice salmon and tofu (for the veggies) dinner for everyone. It was almost easier to wrangle everyone now, because there were so few of them. Andrina and Adella were such big presences that dinners felt so small without them. But, she’d already called them both and told them that she was moving. It had been nice to slide into it that way, but they were also the oldest. She had no idea how the rest of her sisters were going to react.

It was nice to have Panic there for moral support. And her dad, of course. Who was aware that tonight was going to be the night.

Attina was not nervous that they were going to be angry with her. She was actually nervous of the opposite. That people would want her to leave. That they would tell her that why did it matter, why was she making a big deal of this?

She waited until the dinner was winding down. One of the sisters actually got up to put her plate up because Attina was procrastinating.

“Wait—“ she barked, when the sister stood. “Uh, please, sit. I, er—we—“ she glanced at Panic. “Have something to tell you.”

Attina took another breath. “PanicandIaremovingtoAustralia.”

AQUATA: Aquata almost choked on her salmon. “Australia?” she repeated.

This was the thing: Aquata was learning to let go of the idea that all of her sisters would stay in the same place forever. Especially after talking to her at the wedding, Aquata now understood that this wasn’t just a phase for Andy and she probably wasn’t coming back. And even though Adella had gotten the brunt of Aquata’s crisis about the family splitting up, Aquata had eventually made her peace with it.

She maybe expected Tina to leave, eventually, especially since she didn’t have her wedding in Swynlake. But Australia?

She coughed and cleared her throat, her expression going neutral. “That’s a long bloody plane ride.”

ARISTA: Arista chewed on her tofu, happy to just sit among her family. Dinner nights were always so lovely. It was how they all caught up on things. (Well, at least the ones who were still in town).

Her brows shot up, Aquata had a point. It was a long plane ride. “Australia? Like forever?” Australia seemed pretty permanently away from Swynlake. But she trusted her sister to not make decisions unless she had already thought it over like a million times. “You think you’ll be near where Jake’s from? Maybe he can give you some pointers or teach you slang.” She tilted her head, trying to think back to all the weird things she’d heard her sister’s boyfriend say over the years.

ALANA: "Australia?!"

To be honest, Alana had a moment of fear. It struck her suddenly and she looked at the dinner table, at the empty seats where Andrina and Adella once sat. She felt something twist in her gut, but then she looked at Tina and that twisting feeling transformed into a swell of pride.

"Ohmigosh that's so great!" she exclaimed. "Wow, more excuses for me to visit."

MELODY: Well, that was far. Was the first thought as Melody digested the news. She wasn't all that surprised though. Each of the girls were moving on in their own way she wondered who would be next truly.

But this wasn't supposed to be sad. Not at all. "When are you planning on moving?"

PANIC: Panic watched everyone's reactions grimacing ahead of time almost expecting for it to go terribly. Though it wasn't that bad not yet. "It's a long plane ride for sure but we're planning on getting a big enough place you guys can all visit whenever." Panic offered leaving Tina to answer Arista because really who knew especially with school. 

"Toward the end of summer, we need to be there by Late August so we want to settle in a bit."

ATTINA: Attina hadn't announced something like this since...well, since her pregnancy. And she'd done that one on one, because as much as she insisted on these dinners and everyone sharing their lives with each other, she always found it a little overwhelming to be the focus of the attention.

It was nice to have Panic there to help her, because she was trying to untangle her tongue while all her sisters and cousin stared at her. She knew it must be shocking and probably felt like it was coming out of left field and she felt herself about to word vomit everywhere a bunch of excuses.

Panic corralled everyone first and Attina shot him a grateful look before nodding.

"Yes, late summer. I will, uhm, I'm--I accepted an PhD studies offer at the National Australia University in Canberra for Marine Biology. It's not near Jake's, unfortunately." She had already looked that up, because of course she knew where her sister's boyfriend was from. "But, I'm sure he can give us plenty of pointers."

AQUATA: Aquata was stunned.

Andy had always been a loose cannon. Adella had always wanted to get out of Swynlake from the beginning. But Tina? Tina, who basically ran Whosits and Whatsits, who went to every community event, who had a house here and friends and, okay, probably a lot of bad memories too, but who seemed more a part of Swynlake than anyone else?

It was like one of those moments when you were a kid and realized the teacher didn’t sleep at school. Attina had her own life. But.... Australia was really far.

Aquata didn’t really know what to say. She didn’t want to make a scene, or be the unsupportive one (again). But for someone who liked plans and routines, who liked being able to keep track of everyone, it was a lot to process.

“Congrats, Tina, that’s cool,” Aquata said flatly. She needed to go for a swim. She looked at Panic and smirked. Humor was a little easier than being sincere. “Better be a mansion. I’m not getting on a twenty-hour flight for any old cottage. Or maybe I’ll just swim there. Probably faster.”

Aquata made a face to show she was joking, because she didn’t want to freak Attina out, but honestly, it did sound like a fun idea.

ALANA: Alana was still processing this. First Andrina had left, which seemed like a given and honestly, Lana had been surprised it hadn't happened sooner. She missed Andrina so much, her absence like a loose tooth she kept on prodding with her tongue. Adella had left second, which hadn't been a surprise either; she'd already left once the first time and she'd never wanted to stay in Swynlake. She had big dreams and a little town in England didn't exactly lend itself to them.

And now Attina --

Maybe this had been coming from the beginning. Maybe Attina dreamed of things too, dreamed of the ocean and helping save it. Maybe it was obligation that had kept her trapped in Swynlake, that obligation growing into fear and spreading across the rest of them. Maybe that's why Andrina had broken away, why Adella had always dreamed of bigger things. Maybe it all went back to Attina and the fact that she had stayed in the first place, instead of leaving.

Alana always wondered why she was working in a shoppe when she had a Master's degree.

Alana didn't know what she'd do without her big sister. It was one thing for Andrina and Adella to go off -- but Attina?

She wasn't sure where to look. Looking at Attina might make her cry. Looking at Aquata might make her scared. So she looked at Arista, who was soft and smiling and with a deep breath, Lana tried to relax herself.

(She wondered, briefly, if she would ever leave Swynlake; she'd thought about it a lot as a teenager but now... well with her own place and her own life separate from the legacy of her family, it didn't really bother her as much. She wondered, out of all of them, who would go and who would remain).

"Oh my gosh, do you have a new place picked out already?" asked Alana. "Is it by the beach? Please tell me it's by the beach."

ARISTA: Arista, of course, was sad to see any of her sisters go. But she knew they all had to do some exploring of their own some time. She had left for a time to find perspective and where her music came from. Andy had left to grow and spread herself far and wide so she could experience everything the world had to offer. Adella had dreams that were outside of Swynlake. And so while she had been sad for those instances, she was happy for them because she knew they were becoming who they wanted to be most in the world.

And though the sadness over Tina hit her heart particularly hard as Tina had been the most patient and the most kind to her growing up (they of course were not without their fights as seen over the last couple years) but Arista knew that Tina had put a lot on hold and been through so much in town that it only made sense that she too at some point would want to find something different out there. And that didn’t mean they were losing her. They were just gaining distance while she became an even brighter version of herself.

She looked between her sisters, they all has their respective reactions but one thing was clear, they loved their sister and wanted to see her happy in their own ways.

She smiled softly, her voice matched her smile as she looked at her eldest sister, “I think you really deserve this, Teens.”

MELODY: Melody sighed softly but nodded, this was good. The degree was amazing. And it would be good for Attina and Panic, she was sure of it. Or at least she hoped. Just like she hoped all the others girls were happier doing whatever it was they were doing right now.

It felt lonely in a way, seeing everyone move on but making her life more steady here. Would she be like them too one day? Would the rest of the Triton's be like that?

"Congrats Tina. That's going to be such a cool program. " None the less Melody would take each day by day and today was Tina and Panic's day.

PANIC: Panic nodded giving the girls all a smile, a brief glance over to JEFF too. "We'll be near the water and no little cottage don't worry. Though maybe not a mansion big." Panic grinned with a raised eyebrow in Aquata's direction.

"Enough room and good scenery, it'll be worth the flight." Reaching over Panic gave Tina's hand a small squeeze. This was going a lot better than expected and part of him wondered if it was because some of the others girls had left. Strangely enough this was more common than you would first think.

ATTINA: No one was yelling. Everyone was smiling--some more reserved than others. The air was a little stilted, awkward, and nervous, but...no one was yelling. That let Attina know, more than anything else, that this was the right decision. She had let out the breath she was holding and managed a tentative smile herself.

There were little tears shining in the corner of her eyes, but she nodded, squeezing Panic's hand back when he reached for her.

"Yes, of course. You are all welcome to visit, at any time. And we'll all FaceTime and text. It will be like I'm not really gone at all."

"We'll miss you too, Tiny, but don't worry. You're going to do brilliantly," Daddy said, reaching over to squeeze Attina's shoulder. She smiled over at him. "Your mum would be proud."

"Daddy," Attina said, those tears trembling--much more closer to falling. "You must promise to keep me informed of everything. I'll miss you all so much."

[kousin-itt]

I Got Your Back - Part 3

WARNING: Violence and blood in this chapter, along with light implied sexual humor.

Part 3

Block pulled up a holographic screen to display the information as he relayed it to Cavendish and Dakota. “The Triton’s Amulet was a priceless jeweled necklace created by a master jeweler and given to the daughter of a family who was feuding with the jeweler’s family. The feud between the families was full-on bloodshed. I’m talking Montague vs. Capulet level, at each other’s throats at all times. The Triton’s Amulet was a peace offering, a symbol of the friendship between the jeweler and the daughter. It was passed down through the generations until it was donated to a museum, where it was stolen by this man: Tyler Cobalt. Cobalt sells the Triton’s Amulet to a black-market dealer, who later dismantles the necklace and sells the parts for profit. That, gentlemen, is what you are going to prevent.”

“So, we are to prevent the theft of the amulet?” Cavendish guessed.

“Oddly enough, no.” Block said. “See, the Triton’s Amulet being stolen prompts the museum to create better security measures, which later prevents worse thefts from happening. You two are going to tail Cobalt until he steals the amulet, and then you’re going to take the amulet from him before he can sell it.”

“Thieving the thief. I like it.” Dakota chuckled.

“Stay sharp, Chuckles.” Block snapped. “You two won’t have a big window of opportunity. Cobalt steals the amulet late at night and then hops on a plane early the next morning to take the amulet to the black market. Based on our research, Cobalt actually gets killed by the buyer once he gets to the black market; but, ideally, you two dunderheads will retrieve the amulet and Cobalt will be apprehended by the police of his era. Be quick, be careful, and be sneaky.”

Dakota caught the backpack Block tossed to him. Cavendish accepted the paperwork with all their mission information. “We will not let you down, sir.”

“Well, you haven’t so far.” Block shrugged. “But I’m not getting my hopes up. Now get out of here!”

A short while later, the Second-Class Time Travelers were in their time vehicle and speeding off into the time stream.

Dakota fiddled with his seat to get comfortable. “When are we getting our own vehicle? I’m tired of this stupid van.”

“It’s a standard time vehicle for in-training and rookie agents.” Cavendish reasoned. “I’m sure we’ll get a new one once we complete this mission. Besides, it’s not that bad. It’s actually one of the newer models, if I’m not mistaken.”

“Yeah, but the seats could use new stuffing.” Dakota complained.

“If you don’t like it, why don’t you go lay on that mattress you insisted on putting in the back?” Cavendish jerked a thumb behind him. Indeed, the backseats were folded down and there was a twin-size mattress taking up half of the empty space behind them.

“That’s my napping mattress when our missions take too long.” Dakota protested. “If I lay on it now, I’ll fall asleep.”

“I never took you for the type to care.” Cavendish sighed.

“Eh, go boil your head.” Dakota snickered.

“You’re ridiculous.” Cavendish muttered under his breath.

Thankfully, they arrived in the right time and place soon enough. The darkness of the night setting contrasted greatly with the bright colors of the time stream they just exited. Dakota took out the mission file and punched a location into the GPS. Cavendish followed the directions to the museum while Dakota rummaged around in the backpack Block gave them.

“Typical supplies. Rather boring weapons.” Dakota took out two hand pistols, designed appropriately for their current era. Generally, time travelers only used weapons that matched whatever time period they were in, just in case that weapon were to get lost or taken by someone else in that era. Plus, fewer people would ask questions.

“I hope we do not need to use them.” Cavendish was a decent shot, but he despised guns. They had no elegance, no beauty in them. Not to mention it was far too easy for someone to be killed by one.

Dakota hummed in thought. He looked and sounded more focused than Cavendish had seen before. Dakota didn’t like that Block gave them guns. Not because he hated guns, per se, but because he worried about having to use them. Time travelers weren’t supposed to cause deaths in the past unless absolutely necessary.

Cavendish parked the van around the corner from the museum. He and Dakota exited the vehicle and walked up the stairs to the front door of the magnificent building. Dakota pretended to check the hours while covertly investigating the dark interior.

“According to the files, this building’s alarm system only goes off if someone tries to break and enter.” Cavendish said as they walked down the steps.

“But Cobalt gets away and the theft isn’t even discovered until morning.” Dakota remembered that from the file. “How does he get in without tripping the alarm?” He looked ahead and froze, cursing in Italian. Cavendish followed his gaze and saw their target. Cobalt walked towards them, his hands stuffed in his pockets and a cigarette pinched between his lips. He looked inconspicuous, like a barman walking home from a late-night shift.

“Act casual.” Cavendish whispered.

“Does that mean we kiss?” Dakota teased.

Cavendish bit his lip to keep from exploding in indignation and frustration. How can he be joking around at a time like this?! Cavendish wondered. Dakota sniggered again, and the pair kept walking. Cobalt passed them without a second glance.

Smart. Dakota mused. Act like you don’t exist, like you belong where you are now, and most people glance over you. He pulled Cavendish into the next alley they passed, and they watched Cobalt continue down the sidewalk. Once he started climbing the stairs to the museum, Cavendish and Dakota tiptoed closer and hid behind one of the statues out front. They watched in amazement as Cobalt pulled a key from his pocket, unlocked the door to the museum, and strutted inside like he owned the place.

“Clever.” Cavendish tapped his lip in thought as they returned to their van. “Either he stole the key from one of the employees or he found a way to copy it himself.”

“Now we know how he avoided detection until morning.” Dakota said. They sat in their van, moving it only just enough to see the front doors of the museum. “What do you say we jump him when he gets back? Take the amulet, arrest him, and get back home for lunch?”

“Not likely.” Cavendish said. “Did you not see the bulge in his jacket?”

“Was I supposed to see it? What if it’s inappropriate?” Dakota shrugged.

Cavendish face-palmed. Curse his partner’s crude humor. “I’m saying he very clearly had a gun in his pocket. Best if we wait until he comes out. Then, we will follow him home and steal back the Triton’s Amulet while he sleeps. Less chance of us getting shot.”

Dakota checked the time. “Depending on when he gets back and how long it takes to drive there, that only gives us a few hours before his flight in the morning.”

“We can manage.” Cavendish assured.

The confidence in his partner’s voice put Dakota’s mind at ease. They waited, watching the doors until they spotted Cobalt leaving. Cobalt returned to his own car and drove off, with Cavendish and Dakota safely following him. Cobalt left the brightly lit streets and arrived at a dinghy strip of townhouses. Cavendish parked the van in a dark alleyway. Dakota slung their backpack of gear over his shoulder. Cavendish glanced to his satchel in the backseat, and he decided it would be best if he only had his firearm with him. He and Dakota trailed Cobalt’s car to find which building he lived in. Cavendish and Dakota ducked behind some bushes and watched Cobalt park and enter a house. The lights inside came on, and then flipped off fifteen minutes later.

“He’ll probably sleep for a while before his flight.” Dakota guessed. “Let’s get that amulet.”

“Wait a moment. Give him time to really fall asleep.” Cavendish whispered. “If he locked it in a safe, we need time to hack into it.”

“You can crack a safe?” Dakota looked impressed.

“You don’t know everything about me.” Cavendish shrugged with a rather proud smile.

“I hardly know anything about you.” Dakota said. “We really ought to fix that. What’s your favorite food?”

“You really want to do this now?” Cavendish hissed. “What if someone hears us?”

“It’s just past midnight and all the other houses are dark. Relax.” Dakota scoffed.

“I will relax when the mission is complete.” Cavendish declared.

“And then we can talk more? Get to know each other better?” Dakota suggested.

Cavendish opened his mouth to retort, but conflicting words tangled his tongue. Part of him wanted to scoff and assure Dakota that such a thing wasn’t necessary. Another part of him pleaded to say yes. So Cavendish cleared his throat and returned his attention to the house. Despite his mild fear of the dark, he was happy the shadows could mask his discomfort. Why did he find the idea of hanging out with Dakota enjoyable? What was so appealing about the thought that he and this brutish young man could be friends?

Once they agreed it was safe, Cavendish and Dakota snuck up to Cobalt’s house. Dakota picked the lock to the front door, and they crept into the dark house. Dakota handed Cavendish one of the pistols from his backpack. Cavendish gestured for Dakota to go up the stairs, and Dakota nodded to confirm. The pair split off to search the two-level house.

Even in the near-blackness, Cavendish could register the layout of the connected kitchen and living room area. The first word that came to mind was “pathetic.” From the outdated appliances in the kitchen to the weathered easy chair and cheap TV in the living room, Cavendish deduced that Cobalt was a broke loner, which explained why he would steal for profit. If it were different circumstances, Cavendish would entertain the idea of counseling Cobalt instead of leading him to the police. However, Cobalt’s file outlined a history of drug abuse and distribution to high schoolers, so Cavendish felt little pity for the man destined to be arrested that night.

Must focus. Cavendish reminded himself. The sooner Dakota and I find that amulet, the sooner we get out of here.

As Dakota ventured upstairs, Morgan’s lessons echoed in his head. Never step in the middle of the stairs, where creaks are more likely to happen. Keep low to the ground to delay enemy detection. Be aware of everything around you. Listen; do not hear but listen. Dakota felt a tingle go up his spine. He hated silence. He hated the apprehension that came with sneaking around. Any moment now, he might get caught. He wished Cavendish were at his back.

Upstairs, there were three doors, two of which were closed. Snoring came from one door. By reaching into the open door, Dakota could feel a countertop and porcelain sink for a bathroom. Dakota moved deliberately, closer and closer to the closed doors. Since they neighbored each other, Dakota couldn’t tell where the snores came from. After a quick eeny-meeny-miny-moe, Dakota twisted the knob of one door and pushed it open, ever so slowly.

He nearly cursed in Italian again.

Dakota chose the door to the office space, but Cobalt evidently decided to sleep in his rickety office chair before his early flight. Dakota almost shut the door when a glitter caught his eye.

Figures. Dakota silently groaned. The Triton’s Amulet lay on the desk next to Cobalt. Dakota’s mind raced. What would Morgan do? What should he do? What might Cobalt do if Dakota’s plan failed? Dakota had a thought to awaken Cobalt and arrest him, but Block did make it explicitly clear that Cobalt’s fate should rest in the hands of the authorities in this time period. Dakota didn’t much care for the rules. Still, he had no idea if Cobalt had a weapon on him. Better to grab the amulet and get out.

Quietly, Dakota slipped the backpack off his shoulders and set it aside, just in case he had to make a run for it. With his gun trained on Cobalt’s sleeping form, Dakota took the slowest, most silent steps he could manage. He forced his breathing under control to stay quiet. His eyes flickered fast, focusing on Cobalt for a split second and then spending the next half-second staring at the desk so he would not bump into it. Cavendish was still downstairs.

It only took a minute that felt like an hour to reach the desk. Still, Dakota kept his gun pointed at Cobalt. His fingertips gripped the edges of the trident-shaped, jeweled pendent. Dakota lifted the amulet from the desk and slipped it and the chain into his jacket pocket. Dakota moved back to the door, his nerves settling.

He reached the door, and a light cast his shadow against the wall.

Dakota spun around, safety flicked off on his gun, but the brightness of Cobalt’s flashlight blinded him. Cobalt pushed the gun aside and body-slammed Dakota into the wall. He smashed the door against Dakota’s hand and forced him to drop the gun. Dakota shoved Cobalt back and punched him. The flashlight hit Dakota’s face, and he tasted blood. He aimed low and tackled Cobalt like a linebacker, sending them to the ground where they grappled for control. The erratic movement of the light disoriented them. Cobalt may have been similar in build, but he was fueled by desperation, and a desperate man was hard to fight until you found just the right moment to subdue him.

In the poorly lit space, Dakota didn’t see the office chair and tripped over it while trying to escape. The chair landed on top of him, and then so did Cobalt. The flashlight’s beam hit Dakota’s eyes again. The top of the chair’s backrest pressed against his throat, and Cobalt pushed his entire body weight on the chair to keep Dakota pinned.

“Thought you could swipe my prize, huh?” Cobalt sounded as crazy as he looked. “I got someone willing to pay a fortune for that thing, and you are going to pay big time for screwing with me.”

With the chair pressing against his legs, Dakota didn’t have the proper leverage to push or kick it off him. He struggled with all his might, clawing at the poor-quality leather upholstery. He couldn’t get a breath in. His thoughts ran a mile a minute. Where was his gun? Where was Cavendish? What would happen to him if Cobalt realized there was a second intruder in the house?

Dakota fought to stay alive. The room was getting darker. Was the flashlight going out?

“No!”

Cavendish may be lanky in figure, but his full body weight and force of the blow was strong enough to knock Cobalt off Dakota and send both men tumbling across the floor. Dakota coughed harshly, pushing the chair off him. He could just make out Cavendish and Cobalt exchanging punches. Cobalt kept one hand on Cavendish’s gun to keep it away from him. He punched the time traveler in the eye, breaking his glasses. Driven by adrenaline, Cavendish punched back harder. Cobalt managed to get one good hit into Cavendish’s stomach that knocked the wind from him.

Then, Cavendish felt something hard jab him in his ribs. Just before he realized what it was, a gunshot went off, and burning pain coursed through his torso. In shock, Cavendish hit the ground while holding his side. Cobalt had his gun, aimed right at him. Cavendish flinched, preparing himself for another shot that would hopefully kill him immediately.

“Hey!”

Cobalt turned his attention and gun to Dakota. Five shots left Dakota’s gun in quick succession: one hit Cobalt in the forehead, two hit his chest, one hit the wall, and one grazed Cobalt’s shoulder. Cobalt was dead before he hit the floor. Dakota didn’t need to check for a pulse to know that, but he still kicked the gun from the criminal’s hand before he ran to his partner’s side.

[bigskydreaming]

So I’m writing a short story in my Changelings ‘verse for a galaxy brain commission that was simply “Boyd gets a boyfriend.” Nothing wrong there. But it does mean I feel like I finally gotta nail down the full intricacies of Boyd’s magic, which is unnecessarily....extra, due to the existence of the Curse of the Specialist Snowflake that afflicts me. 

Like, I couldn’t just give him one physical change and one magic like most other Changelings, nooooooo, he’s gotta be the Zodiac Kid and have twelve completely different changes and magics, one for each sign and he changes throughout the year in accordance to the changing of the signs. Why’d I do it that way? Cuz I gotta, obviously. Hahahaha, shut up, me.

I mean, there was a point to it originally, like, his sister Alicia’s Change and magic is that she’s surrounded by ghostly versions of a bunch of her alternate universe selves at any given moment, and she can communicate with them and call upon their skills and is sometimes called Sibyl around town, so like, the common thread/theme of her and Boyd’s magic is like, an infinity of self, but also, by “there was a point to it originally” I also just mean I couldn’t settle on and pick what I wanted Boyd’s magic to be so I was like here’s a fun thing I could do, make up a reason for him to have even MORE options.

.....I feel like, potentially, this self-created problem I’m having could have been avoided somehow, but gosh, fuck if I can see how.

So most of Boyd’s magic is hammered down....

In his Sagittarius phase, he’s physically a centaur whose lower half looks like a night sky studded through with starfields and moving constellations, and his magic from November 23rd to December 21st is uncanny intuition, a kind of off-the-top-of-his-head divination. He just instinctively knows the answers to questions when he’s in his Sagittarius phase, the more specific the question the more accurate his answer. He doesn’t have context, he doesn’t KNOW how he knows the things that he knows, he just knows them. 

You ask him if you’ll meet someone who could very plausibly be the love of your life within the next week, and if he says yes, its absolutely, one hundred percent true and accurate.....but if you ask him then how can I find them, he might say well first thing you gotta do is get out of my face and stop bugging me, and then walk down the street and turn left.....and that too, would be absolutely true and accurately point you in the direction that answers your question but like, that’s all he’s got and you’re missing a few steps at that point. So. Its not an exact science or anything. 

There’s definitely tricks to maximizing the intel you get out of him when he feels like being hit up for intel and doesn’t just string you along to be a total troll. Which he is. Hence why he also spends most of December filling in all of Alicia’s large stockpile of crosswords and sudokus, as he just instinctively and whimsically knows what to fill in even if he doesn’t actually have a clue WHY the Mesozoic Era is the answer to 49 Down. Alicia maintains this is cheating. He maintains he doesn’t care, and answers 64 across with “Johannesburg.” Fuck if he knows why.

In his Capricorn phase, he sublets his apartment in Bordertown and takes a month lease on a place underwater in Sub Francisco.....as for this period he’s more of a classical merman, but also if a classical merman has goat horns. But like, whose to say they don’t? Have you ever met one? Thought so. His magic during this period is horns of plenty.....he touches something to one of his horns, and comes away holding two of that item. So it basically only works on things that can be hand-held, but other than that it could be food, physical items, medicine, etc.

In his Aquarius phase, he’s physically the same as he was when a normal mainstream human....except that his skin has a faintly translucent quality as if that of water, with faint stars spiraling in distant patterns in the depths of his slightly watery form. He’s not invisible or actually translucent, like you can’t see through him or even see his organs or anything....rather, its more like most of the time he just looks like normal, human Boyd, albeit wearing a lot of glitter for some reason....but then he turns and catches the light just so, and for a moment you can see how the light is shining THROUGH him similar to the way it shines through the surface of a body of water. His magic in this phase is based on Ganymede, the cup-bearer of the gods.....water he touches becomes imbued with healing properties. 

Like, he unscrews an ordinary bottle of Evian and reaches in with the tip of his finger, and whammo, you’ve got a healing potion whose magic will last until the last of the water is gone, even if its not used until after his Aquarius phase. Once he’s imbued a source of water with his magic, its a healing potion from then on out, which makes his shop a useful place to find healing magic year-round.....or at least until he runs out. You come into his shop even in late July, if he’s still got some healing potions lying around they’re as good as ever....but if he’s already out of stock, you’re out of luck until next year’s Aquarius phase rolls around, and you gotta go look elsewhere on Mercy Row for a healer in the meanwhile.

In his Pisces phase, he goes back to his Sub Francisco apartment as his physical Change here is that of a centaur-triton....kinda the classic ithyocentaur, but also he’s got kinda webbed-like wings spanning from his back.....its a whole thing. But a good look. His magic in this phase is ‘the perfect pair’....he cups his hands and forms a magical, elemental construct in the shape of a koi fish that’ll swim off in the direction of the perfect complement to something or someone. Like a personal, magical compass guiding him or whomever he makes one for to like, the magnetic north that exists somewhere as the perfect counterpart to an already present south pole. Whatever, this makes no sense and is super confusing, I get that. But it makes perfect sense and is not confusing in my head. Still just working on translating from point Head to point Paper without the concept getting lost in the translation.

In his Aries phase, he’s his ‘normal’ mainstream-appearing base form....but also he’s got golden wings and ram horns. And his magic is that he breathes fire. But its magic fire, and it doesn’t burn things it touches, it turns them to gold. Its a whole Golden Fleece thing mixed with a King Midas thing.

In his Taurus phase.....idk yet. I mean, bull horns seems a pretty obvious go-to motif, but haven’t decided for his magic yet. Mostly because @sunwukxng is a Taurus and he takes things like this juuuuuuuust personal enough that I know if I don’t come up with something good here he’ll decide its a mortal insult towards all Tauri and thus him as well, and he’ll declare a blood feud against me that shall last five hundred years. And he’s already got like, six of those in progress, so....trying to avoid racking up anymore.

In his Gemini phase, he splits into two selves, his physical self and astral self, so he’s basically followed everywhere by both his shadow and a spirit version of himself he can astral project elsewhere. And when his astral self goes through something, kinda like how there’s that idea that when a ghost passes through someone they feel a chill or as if they almost left their body for a second.....his astral self can kinda like....yoink other peoples’ astral selves out of their physical forms temporarily. Yoink is of course the highly technical, scientific term for the phenomenon. Obviously.

In his Cancer phase.....idk yet. Look, its hard to be classy and deep concept-y about crabs, okay? I’m sorry Cancers, but even in the original myths about the constellation the gods were like yeah, this crab constellation was a mistake, we just felt bad about Artemis kicking that one all the way up into the heavens after it pinched Orion, the only himbo she ever almost made a boytoy out of. A lot of other Zodiacs featured animals that had the common perception of being viewed as pushing the sun across the sky, so there’s maybe something there in that direction....idk yet.

In his Leo phase, Boyd mostly appears as his ‘normal’ base mainstream-human looking self....except he’s big in a way that is not exactly Giant Big (and there are a good couple dozen giant-sized Changelings living in Bordertown, so like, he isn’t officially anywhere near that) but he is possessed of a height and size that are nevertheless best described as Ridiculous. Like he’s bigger than the Strange Angel, but not as big as Teddy the Sun-snake. That means nothing to anyone but me. Whatever. Moving on. Anyway, in his Leo phase he also has a fiery red mane kinda....that’s threaded through with silver stars in the Leo constellation shape. His magic in this phase is his ‘roar of command’....basically, he can imbue anything he says with the force of his will, and thus like.....make inanimate objects do what he tells them. He’s like “Open sesame” and a locked door is like you got it, boss.

In his Virgo phase.....I am not entirely sure what he looks like yet. I’m pretty sure its mostly his mainstream-looking ‘base self’ but like.....with an unnatural charisma, but also something a little less generic than that. Needs a specific hook still, beyond just “I’m ridick pretty.” Like, he’s Sinqua Walls. He’s always ridick pretty. Its a thing. But his magic in this phase is like.....an aura of inspiration. He just sorta puts out a vibe that inspires people around him in various ways that are unique to the individual. 

He’s also way shorter than he used to be even before his Change hit, or like, shorter than he is in any of his other phases. Like, we’re talking five and a half feet max. Is there any reason for this whatsoever beyond me being amused at the juxtaposition of him going from his Biggest and Most Ridiculous Phase immediately to his Smallest and Most Ridiculous Phase? No. No there is not. Fear the deep profundity of my creative process, for it is deep and it tis profound.

In his Libra phase....I’m not sure yet what his physical change is, but I do know his skin has a scaled texture, and I know those aren’t the scales normally associated with Libra but there’s a point to it, its just......missing some ingredients still to totally make sense. But he also tends to wear a blindfold a lot of the time during his Libra phase because I know his magic here has to do with viewing things in terms of overlapping possibilities and potentialities, like, he looks at things and sees not just the way they ARE but also a whole bunch of different ways they COULD be, and his magic enables him to kinda tweak things from the way they are more towards one of those other possibilities, in a sense that’s all about ‘balancing’ things.

However, the sensory stimuli of all those potentialities 24/7 is honestly overwhelming and gives him serious migraines, so he mostly tries to rest his eyes behind some kind of blindfold most of the time and just take it off to focus on specific things in order to utilize his magic, and only a little at a time. This is probably the most technically ‘powerful’ of his magics in terms of both versatility and scope, but it takes a toll and he really tries to pace himself during his Libra phase or it just gets to be too much, really fast.

And then lastly, in his Scorpio phase, he’s got a whole scorpion tail and barbed stingers along the outsides of his arms.....but rather than lean into the whole ‘eww Scorpios are duplicitous and manipulative’ and also ‘eww scorpions,’ his magic here calls back to how scorpions are actually protective guardians in a lot of mythologies and folklore, and there’s also plenty of Zodiac lore about the idea of the Scorpio sign being all about uncovering truths at any cost, etc. So his magic here is another ‘liquid/mutable’ type and takes the form of venom from his stingers that’s almost like drops of amber.....and that have various properties aimed at uncovering truths and exposing lies and falsehoods. Like, elixirs made during his Scorpio phase are another highly sought ware at his shop, because depending on the venom concentration he places in each, they have different properties. Like, he’s got vials where just a drop placed in your drink or on your food will reveal if its been drugged or poisoned, or vials of elixirs that basically act as magical truth serum.....and then other vials where like, you splash a little of it on something you suspect to be an illusion or a magical disguise of some kind, and it’ll dispel the false magic and reveal what’s truly there. 

“What happens if like, I splash it in the face of someone I think is wearing an illusion disguise and it turns out they’re not and that’s really just what they look like?” A customer asks.

Boyd shrugs. “Then they’ll just be wet. And probably a little pissed, I imagine. That’ll be $49.95 please.”

“I thought you said it was $29.95!”

“That was before the Asked Stupid Questions tax was applied,” Boyd smiles placidly. “Will that be cash or credit?”

 Okay. So. Typing that all out cleared up some of that for me but not quite as much as I was hoping as Cancer and Taurus apparently still persist in Vexing me.

Hmmmm.

[sheenaghsmusicblog-blog]

Music Theory Blog by Sheenagh Shannon (MK)

My name is Sheenagh Shannon. I’m an 18-year-old singer/songwriter currently studying music performance. In this blog I’m going to break down the basic principles of music theory to hopefully help any readers gain a better understanding of the inner-workings of the music we love and listen to.

Scales  

I’d like to begin by explaining what scales are and how they work. The pictures I will be showing have music written on what is called a stave, and included on the stave is either a bass or treble clef (treble clef being higher, and bass clef being lower.) The notes in the treble clef start with E on the bottom line and ascend from there, whereas the notes in the bass clef start with B and ascend from there.

(Left is the bass clef, right is treble clef.)  

A scale is a group of notes that are arranged by ascending or descending order of pitch. In an ascending scale, each note is higher in pitch than the last one and in a descending scale, each note is lower in pitch than the last one. I will explore the two most common types of scales; major and minor.

You must have a note on every single line or space of the stave. One of the most common types of scale is the major scale. These scales are used in an endless variety of songs we listen to. Major scales are defined by the order of which semitones and tones occur. Semitones are whole steps and half steps, and their combination specifies how a scale should be interpreted. For example, the major scale is this:

Tone – Tone – Semitone – Tone – Tone – Tone – Semitone

Or in whole steps and half steps it would look like:  

Whole – Whole – Half – Whole – Whole – Whole – Half

This formula can be used to form a major scale starting on any note. Major keys are often correlated with positive emotions – a good example of how a major key can create an upbeat and joyous mood is a song called ‘Breakfast At Tiffany’s,’ by Deep Blue Something. (https://www.youtube.com/watch?v=QSgJ5On8Zso)

The second type of scale I’d like to explain is the minor scale. A minor scale, much like the major scale, also has seven notes but is defined by having a flattened third. This means that the third note on the scales is three semitones above the first note. With major scales, the third note of the scale is situated four semitones above. A seemingly small change, but it makes all the difference!

Tone – Semitone – Tone – Tone – Semitone – Tone – Tone

It’s important to note that there are three different types of minor scale: the natural minor, the harmonic minor, and the melodic minor. Each type of minor scale uses an ever so slightly different formula of semitones and tones, but they all share the minor third in common.  

Minor keys are more often associated with music that feels sad or melancholy, but they don’t necessarily have to abide by their assigned moods – minor keys can create intensity, passion, or with the right placement, even joy. Linked is a song called ‘Losing My Religion’ by R.E.M  (https://www.youtube.com/watch?v=OKvCV8MFIaw) a song that begins on the chord of A minor. I believe this is a good example of what a ‘minor’ song sounds like.  

Modes  

Next, I’d like to discuss modes. Modes are a series of seven musical scales each with definitive qualities and varying sounds. Their names are:

Ionian  

Dorian  

Phrygian  

Lydian  

Mixolydian

Aeolian

Locrian  

Each of these scales are a type of diatonic scale, which means they have seven notes and have two intervals that are semitones (half steps) and five intervals that are tones (whole steps.) The modes can also be split into two types, major and minor modes. The three major modes are Lydian, Ionian and Mixolydian and the four minor modes are Dorian, Aeolian, Phrygian and Locrian, although it is important to note that the Locrian mode is typically defined as a diminished scale. I’m going to give a couple of examples to show how modes work.

Ionian Mode

As shown above, the Ionian mode is exactly the same as the C major scale with no sharps or flats. It’s just another name for it!

Mixolydian Mode

The only difference between this mode and the Ionian mode is that it has a flattened 7th note as shown above. A song that uses this mode is a song called ‘Wicked Game’ by Chris Isaak (https://www.youtube.com/watch?v=aid2vMbCNP8) a song that I personally love and listen to quite often. The song is characterized by its haunting and sorrowfully conflicted tone, which exemplifies how modes can be used to create very specific moods or feelings.

Lydian Mode

The last mode I’m going to show you is called the Lydian mode. It’s the brightest sounding of the modes - quite like the Ionian mode, except it contains a raised fourth note. To play a Lydian scale we sharpen the fourth note of the scale. A song that uses this mode is a song called ‘Flying in a Blue Dream’ by Joe Satriani. (https://www.youtube.com/watch?v=SINl5JY7LhI)

Chords  

Chords are the vertical arrangement of notes from a scale. Many people define chords as several notes played simultaneously. Let’s begin with examples of the most basic chord - the triad.

The triad is a class of chords, specifically three-note chords formed by this formula: 1-3-5 or root, third, fifth. There are four kinds:

The major is consonant; the minor is less consonant, but still largely so on most occasions. The augmented chord is very dissonant, and the diminished chord is incredibly dissonant because it contains a tritone (an augmented fourth, or on this occasion, diminished fifth.) This is the basic principle of how chords are formed and what they are. Most songs in existence are built upon a series of chords.

Triads are often extended with some form of the seventh to form a seventh chord. When a chord is called a seventh chord, usually the dominant seventh is meant, which is a major triad with a minor seventh.  Because all sevenths are dissonant intervals, any seventh chord is dissonant, because all sevenths are dissonant intervals. A seventh chord is far tenser than a major or minor chord. Jazz music, for example, is a genre that uses seventh chords quite consistently and often seems to treat them as consonant.  

Keys

In music, the key identifies the tonal centre of a song. This tonal centre is a note that the whole song revolves around, and each note in the song gravitates towards that home base note. If a song is in the key of C, then each note in the song gravitates towards a C. Below is a list of the 12 keys of music. Key of C Key of Db / C# (en-harmonic keys) Key of D Key of Eb Key of E Key of F Key of Gb / Key of F# (en-harmonic keys) Key of G Key of Ab Key of A Key of Bb Key of B / Key of Cb (en-harmonic keys)

Key signatures can provide an idea as to what key a song is written in. Key signatures appear directly after the clef signs and will tell you what sharps or flats will appear in a song. The number of sharps or flats in the key signature can give you a clue as to what key a song is in. For example, if a song has no sharps or flats in the key signature, the song may be in the key of C. If a song has one sharp in the key signature, it may be in the key of G. I’ve included a chart of key signatures to help explain this principle better.

Note Lengths  

Each note written on the stave has a duration as well as pitch. In music notation, a note value indicates the duration of the note. It is the design of the note that tells you its duration, in the same way as the position on the staff tells you the pitch.

This chart should indicate a clear and simple analysis of how note lengths work. For example, if a song is in 4/4, it would look something like this:

Time Signatures

In music, a time signature alerts you to the meter of the piece that you’re playing. The two numbers in the time signature tell you how many beats are in each measure of music. A piece with a time signature of 4/4 has four quarter note beats; each measure with a 3/4 meter has three quarter note beats; and each measure of 2/4 time has two quarter note beats. The most common time signatures you will come across are:

4/4 - Common time

3/4 - Waltz time

2/4 - March time

6/8 time

The most common meter in music is 4/4. In 4/4, the stacked numbers tell you that each measure contains four quarter note beats. So, to count 4/4 meter, if you tap the beat each time, you would be tapping the equivalent of one quarter note. ¾ is the second most common meter, and each measure has three quarter note beats. In 3/4 time, beat 1 of each measure is the downbeat, and beats 2 and 3 are the upbeats. It’s quite common amongst country music.

Once you start listening closely, you can hear the difference between time signatures. A famous song that uses 4/4 is a song called ‘Runaway Train’ by Soul Asylum (https://www.youtube.com/watch?v=OzzKBxlIiEQ.) If you count the beats you can hear the difference between that song and a song like ‘Strawberry Wine’ by Deana Carter (https://www.youtube.com/watch?v=tdWV7PpeTVA) for example.

My Transcription

I’ve transcribed a verse and a chorus from a song called ‘Molly’s Lips,’ by Nirvana (https://www.youtube.com/watch?v=f76GsOBxUg0) 

I’ve chosen this song because it’s simple, to be perfectly honest! It gives a good, clear example of what a transcription looks like. 

All References And Sources Used:

https://en.wikibooks.org/wiki/Music_Theory/Chords

http://www.zebrakeys.com/lessons/beginner/musictheory/?id=12

https://www.uberchord.com/blog/music-theory-chords/

https://www.dummies.com/art-center/music/piano/common-music-time-signatures/

https://www.libertyparkmusic.com/musical-time-signatures/

https://www.essential-music-theory.com/music-note.html

I hope reading this blog has been as informative as it has been interesting. Until next time!

Sheenagh :)

[punishandenslavesuckers]

Somewhere in Faerûn, there’s a tabaxi, a bugbear, a werewolf, and a tiefling all sitting in a boat. There’s a triton in the water but she’s not alone. In a split second, one of the others will have to do something about it. An excerpt from my last homegame session. Combat-heavy one-shot.

Blue and Will are and have been giving each other the silent treatment for the better part of six hours now.

Rime is professionally friendly, but he can only make so much neutral single party conversation before getting annoyed and settling in to watch the river. Their three-boat caravan of small outrigger canoes continues steadily up river – powered by the tireless efforts of the hunkered bugbear jammed uncomfortably into the lead boat. With him is Bian – their smallish tabaxi navigator who’s perched somewhat absurdly on the back tip of the canoe like a lightly armored counter balance.

The second boat, lashed between Rime and Bian’s respectively, is empty save a single occupant. He lies very still, shivering occasionally beneath the worn travel blanket that Rime very carefully tucked around him some hours ago. Tivas hasn’t regained consciousness since the closing of the water purification ritual that went non-stop these last thirty-six hours. Rime was careful to pack the ritual instrument – the Blossom of Beauties – into the druid’s pack and tuck it protectively under his arm.

Tivas, even thralled by delirium, pulled the sacred thing close (pulled close the vehicle of his death) and Rime had to get back into the third boat. Blue commented, eventually, that the flowers ringing Rime’s headband were a weird silver and he told her, simply, “Yeah they do that,” with no further explanation.

So Rime is still in the back boat when they reach the salt marsh.

Bian has her back to Rime so he can see the twitching white length of her tail going this way and that as she scans the foggy western shore. Occasionally, she shoots Rime meaningful looks and Rime grimaces significantly back. Vorgut, the big black-furred bugbear, rows furiously while likewise sending glances toward the reed-choked river bank. His giant, tattered, bat-like ears swivel nervously.

Somewhere to Rime’s right, Blue rows as well, but less out of geographical anxiety than pure, domestic rage and need to put that rage somewhere. Preferably in the water and not directly into the back of her husband’s half-elf skull. She’s a small, blue and white blur of flexing arm muscle, muttering softly to herself in furious Aquan. So she doesn’t notice Rime taking a more attentive crouch in the boat.

Will looks up from his book and scans the waters.

“Hey, Rime.”

“Mmm,” says Rime, rather than use thaumaturgy to speak just yet.

“What’s going on? I’ve got the heebie-jeeebies.”

Rime snaps his fingers and the spell murmurs almost directly into Will’s ear: “Shh. Lizard folk live out there.” He jerks his head toward the salt marsh. “We need to be careful.”

“They wanna eat us,” Bian says without looking away from what she’s doing up front.

Blue, hearing this, growls something like, “Good,” under her breath.

And, naturally, that’s when the first javelin slams with a loud thunk into the side of Tivas’ canoe. The entire party stares, horrified, for moment. Except of course for Blue, who lunges eagerly to her feet with her wand in hand and anticipating a target. That does not happen because a second javelin already airborne immediately slams into Blue’s stomach with such force it knocks her with a scream into the bottom of the boat and blood splatters across Rime’s startled face.

Will howls, “Blue!”

But the javelins have already begun to rain down.

“Bian!” Rime hooks his arm through his battle shield and lunges back toward Blue. “Get us out of here!”

Blue – teeth bared, screaming like a banshee – is snapping the javelin between her webbed fists. She hurls the long part of the shaft into the water, keeping the head of the spear embedded in her gut. She doesn’t waver or try to rise. She raises her hands and instantly, an unnatural slither of white mist condenses from the river waters. A nearly opaque cloud-wall forms between the shore and their boat, veiling them in a literal smokescreen. Rime feels a wind driving like a kick into the back of the third canoe and Blue just lies there, bright-eyed and snarling, her off-hand gripping the javelin.

Magic floods Rime’s hands.

It courses hot from his heart down the track of his arms to his palms. He wills the magic like lightning courses to ground – Blue, Will, and Bian. The blessing diffuses through each of them. Unfortunately, it happens precisely as Will is attempting to jump from their boat into the middle boat and he nearly biffs it, boot slipping so he topples head first into Tivas’ canoe. Bian is already yelling to Vorgut to row faster and easterly. The boats begin to swing toward the opposite shore, far away from the marsh.

Javelins soar from the fog – hitting the walls of the boats, the water around them. One nearly wings the ropes that lash the third boat to the second. Seeing this, Blue waves a hand and a ripple-like mirage passes over the ropes… then a knotted tangle of a dozen ropes appears there, obscuring the target. Will, meanwhile, wrenches a javelin from the side of the boat and without warning, he swells. His spine bows up. Dark fur erupts from the back of his neck and spreads instantly across his body and as Rime watches, stunned, the newly shifted werewolf winds one massive arm back and whips the javelin right back across the shore.

Then he does it again. Over and over. Across the shore, there’s a scream as a spear going ninety miles per hour surely smashes through several lizard men.

Rime maneuvers to the back of the boat and kneels directly between Blue and the foggy enemy-infested shore. They crouch together behind his shield, peering into the mist but no more javelins come. There’s just… motion somewhere in the long grasses and reeds. As Rime’s vision adjusts to the fog… he realizes the shore is literally swarming with lizard folk. The reeds bristle with spears and glinting eyes.

Rime braces the shield more securely. “You see them?”

“Oh, yessss I do,” Blue hisses.

Rime hears her flick the wand of magic missile somewhere directly over his head. There’s a flash and eighteen screaming beads of neon light rip across the river like tiny, hyper-speed fireflies before arching up, then divebombing into the crowd. There’s a sound of wet screaming and bursting. Gore and skull fragments pop as if from red balloons along the shore. Behind him, Rime hears Blue muttering in Aquan and he’d bet it means, “Fuck you fuck you fuck you, I win!”

Will, seeing his wife at work, lays down his sword a moment and grabs the oars to join Vorgut in furious rowing.

Rime – seeing this and hearing the continued guttural shrieks of torment still issuing from Blue’s blast zone – closes his eyes. He presses one hand to his chest, over the three-star sigil of Lliira and for a moment simply mouths, “Show me,” and looks skyward.

It’s instantaneous. The knowing rushes through his head and through his body, takes possession of him in a jolt of sudden muscle memory. Rime shudders, then grabs the oars from where Blue left them and with a sailor’s stolen confidence, begins to pull them asymmetrically through the waters, swinging the tail boat into Will’s rowing, and then into Vorgut’s.

And the boats are suddenly traveling snug to the western shore, so far beyond the range of the javelins that again, no weapons are thrown. For a full ten minutes, Rime expertly navigates the outrigger along the edge of the shore until the light of Lliira fades like a touch from his mind and he loses that sailor’s expertise easy as amnesia. At the front of the boat, Bian is alertly watching the river with one eye and Vorgut’s navigation with the other.

Will, in his boat, says, “Fuck. They’re coming.”

There are splashes from the far shore. Bodies getting quickly through the waters toward them as about forty lizard folk abandon long-range in favor of swimming directly at their small canoes. Blue, behind Rime, staggers up still impaled by the javelin. She hisses, “Let me at ‘em!” and before Rime can tell her to stop fucking moving with a spear in her gut, she raises her arcane focus and throws a fistful of sand into the water.

Magic flashes. Suddenly about half the charging lizard folk go limp mid-swim. Rime sees their eyes slide peacefully, magically shut as they are sucked down by the river’s current and disappear beneath the dark waters.

A force of over twenty furious, screaming lizard-folk are still powering like scaly, ravenous missiles through the water. Rime again takes position between Blue and the enemy, pulling her close behind him and bringing the shield up in one hand. With the other, he raises it palm out toward the waters… and he hesitates.

He can hear Will yelling and hacking furiously as the first wave of lizard men attempt to swarm his canoe. Bian is hissing and snarling, just beheading and hacking into the water. Rime can smell Blue’s blood on the wood and slick on her dress.

Lizard folk hit the third boat.

Wounded, the water frothing with blood, they claw and grab. Gored by Bian, mutilated by Will, the survivors bump down the line of the outriggers to claw madly at the last boat. The grapple the rigs, pulling themselves up, trying to get at both Rime and the wounded sorceress behind him only to be bashed in the face by a shield, but they’re starting to pile on. Rime can’t… he can’t just drive them off. They dragg the boat like an anchor, water sloshing into the…

“Fuck,” Rime whispers.

And summons his spiritual weapon.

It manifests instantly, a bright spinning ball of carnival ribbons hovering like a giant dandelion tuft just above the water… then it swings down, gliding to skim the water, the ribbons foaming the surface as it hooks down to pass along the right side of the head boat where, in a spray of pureed bone, blood, and meat, the razor-sharp ribbons shear one lizard man’s arm off at the elbow, then beheads the fellow behind him. Then on down the line like a meteorite of frothing water and blood, dismembering and bludgeoning any clinging enemies until it reaches that last boat.

The weapon stops directly in front of Rime and grinds a bloody, screaming, person-free space into the waters next to Rime’s shield. This does not last. There are… far too many and even the horror and losses don’t seem to sway them. The lizard folk bash against Rime’s still raised shield, hooked over the side of the boat and he shoves them again, bashes one of them in the face, watches that face shred off the skull when he falls into the weapon.

Blue grabs Rime’s shoulder.

She hisses a little frantically in his ear, “How do you feel about taking a hit?”

To which Rime grits, cheerfully, using his real, demonic voice in all its hissing horror, “Pretty good!”

“Okay!”

Then Blue dives off the back of the boat like a suicidal swimmer toward the lizard-folk infested waters. She arcs up, twisting midair, arms out. A wind catches her Triton frame like a slender kite on an updraft. It carries her upwards, spinning her so for a strange, impossible moment she is almost vertical, upside-down, white hair blown out around her face with her hand out… and she casts thunder wave.

She casts directly at the last boat, Rime, and all the lizard folk upon it.

Rime slams his shield down, grabs a bench and braces as the electricity hits him in a white-hot, screaming wave of pain. Every muscle seizes with a hideous rigor as the lightning courses through him. It hits him like a blow to blast him back, but he holds the fuck onto the boat. He hears the wood splinter, water flood over his boots. For a horrible moment he sees white, then stars, and then the sky reeling above him. Lizard folk are still screaming. He can hear them scrabbling at the out rigging, banging into the canoe walls as they still, still keep on coming.

Then Blue drops with a little shriek into the boat again, almost knocking Rime over, and Will is bellowing, “Jump! Jump! You gotta get in here!”

Rime moves on a dizzy, static-buzzed instinct. He rolls, pivoting to face the middle boat and Will who stands at the back with one hand frantically out stretched and the other holding a blazing scimitar. Rime staggers, still seeing stars and feeling the buzz in his bones. He reaches the front of the boat, drives one foot down on the bow and jumps, landing directly in the second boat and immediately losing his shield from his static-numb arm.

Blue. Is Blue –?

He turns… just in time to see Blue try to follow him. He’s in time to watch it happen, as if in slow motion, as the sorceress’ blood-slick boot squeaks out from beneath the driving lunge off her right leg… and she loses half of her momentum instantly. Rime watches her fall, one arm outstretched toward them – Will and Rime both staring in horror – as she falls into the writhing, blood-red waters.

Split second: Will is screaming.

Split second: The lizard folk start to swarm.

Split second: Bian yells.

And Rime feels his focus like a razor’s edge along the arcane line from his mind to the spiritual weapon. The cyclone ball of ribbon rockets up the side of the boat and every ribbon in its composition loses any bluntness they formerly possessed. What hits the bodies of the lizard folk hits with molecular-sharp indifference and with no clear difference in texture between bone, meat, and water, the weapon plows down into the river and the waves blacken, then redden, then thicken with blood and body parts. The ribbons are no longer any other color but blood red.

Blue tucks into a ball as the weapon screams a horrifying orbit around her, over and over, clearing a ring of mutilation. It’s so precise, it never touches her. Only the ones it intends to harm.

Eventually, the river is clear around her. The remaining lizard folk still alive, screaming, and mauled, swim away back toward the marsh. Blue unfolds herself beneath the water, kicks up, and her head breaks the water by the back of the second canoe now cut free of the third. The water is red around her pale blue face. She blinks up at Rime, bright eyes a little shell-shocked and glassy. Her white hair is pink as Rime catches her arm with a shaking hand and pulls her from the river.

Rime does it carefully, turning her onto her back as he drags her into the bed of the boat.

Will is already pulling Tivas (still unconscious) to the other side of the boat to make room as Rime arranges the tiny Triton woman on the floor and immediately checks the fucking head of the javelin still stuck between her hip bone and her belly. Rime promptly rips her dress open a little to get a better look and presses his fingers into the flesh around the puncture, trying to gauge the depth. Blood pulses around the head of the spear, turning the water puddled beneath her a dark red.

“Blue.” Rime smacks her cheek a little until she looks at him, his real voice grating and seething with whispers. “Are you okay?”

“Eh-heh?” Blue kind of whines.

Rime takes that to mean she’s in shock and therefore not fully aware. So with one hand he yanks the javelin from her gut and with the over dumps a minty-hot rush of healing magic down the tunnel of shredded muscle and perforated gut. Lightly perforated though. Just barely. Rime feels the magic knit her back together until the spell runs dry beneath his fingers. He peels his blood-tacky palm from her stomach… and there’s nothing but a shallow, scabbing cut where a four-inch gouge once gaped.

“You’ll gonna be fine,” Rime says, water dripping from the circlet of flowers around his head. He knows without seeing them, they will be an eye-aching orange. “Dummy. Why the hell did you stand up? You just wanted to give them a target? Give me something to do?”

Blue kinda grins dimly up at him.

“I got ‘em,” she slurs.

Rime looks over the side of the boat, to the receding red waters as the river current eddies and pulls the blood out to sea. He thinks, vaguely, I’ve just lost count. I don’t know how many I’ve killed now. Then he looks back the Blue and Will and Bian perched anxiously at the back of the first boat. Bian’s eyes are big, her tail fluffed anxiously out. He puts on a smile.

“We’re fine,” he says. “Let’s get to Daggerford.”

[royalcordelia]

Time Turns to Amber (2/11)

Summary: The line between universes is blurred when Anne Shirley of Green Gables suddenly switches lives with Ann Shirley-Cuthbert, a university student living in the contemporary world. Suddenly Anne must learn how to navigate the modern world, one which contains a boyfriend, a part time job, and another year of university. Meanwhile, Ann struggles to tackle corsets, farming, and a world without electricity. Maybe it wouldn’t be so bad, but most people can’t tell the difference between the redhead they know and the girl who replaced her. Dedicated to the ever-beautiful @hecksinki

A Time Travel, Soulmate AU

Rated T+ • 7k words • Read on ao3 • Part 1 •

If the circumstances had been different, Ann might’ve secretly wished to never move from this spot. Where else was better than this corner of the world with its enduringly beautiful Avonlea sunsets and comfortable July breeze? She also didn’t mind leaning up against the armrest of the plush porch swing with her legs on Gilbert’s lap. Her eyes fell on him and the one hand that rested on her leg, the other typing something into his phone.

“Alright, Ann-girl. I’ve successfully hacked into your bluetooth speakers. The music choice is yours,” he said. His fingers ran up the skin of her leg in a particularly gentle caress, sending chills down her nerves.

“Gilbert Blythe letting someone else choose the music for once? The world must be ending.”

“Ha ha,” he replied sarcastically. “Maybe I’ll blast something raunchy and obscene so that even Rachel Lynde blushes two miles away.”

“You wouldn’t ,” Ann gasped. Temptation slipped into his face, but drained away as quickly as it came.

“Not today I wouldn’t,” Gilbert admitted. Not with Matthew in the hospital in the middle of an open heart surgery after a small stroke he’d had in the fields early the day before. With Matthew condemned to the ICU, Marilla was glued to his side. She insisted that Ann stay home, an assertion that the 22-year-old redhead rebutted with a fierce conviction. But then Marilla began to cry, and Ann realized that the cost of this battle was more than she was willing to pay.

She called Gilbert, who arrived almost instantaneously to drive her home, and the rest was history - less than a day of moving through the house with the ghost of Matthew following her and the eminence of the inevitable looming over her head. The only thing that kept her eyes from glazing over completely was Gilbert’s kind presence at her side - humble and empathetic. What would she do without him, her very best friend who cooked her comfort food and held her when she felt she’d drown in worry?

He was one of the only people who understood her. He was the only one that could have known that when her eyes burned from so many tears that the cure was the spirit of the island in its sunset and summer wind. Only Gilbert could have known that the one place she could rest was on the veranda of her home, swaying on her favorite porch swing and listening to her favorite music.

“Hey, where have you wandered off to?” Gilbert asked gently, scratching his fingers into the skin behind her ear. “You’ve been staring at Hozier’s album cover for a good minute now.”

“What can I say? I love to appreciate art,” she replied weakly, pressed play, then handed Gilbert his phone. As the opening notes of “In a Week” hummed from the small speaker set on the porch railing, Ann shifted so that her head was buried in his neck and his arms could wrap around her like a protective shield.

Yes, if circumstances had been different, she’d be running away from her rapid heartbeat and the peace of being the recipient of many head kisses. And Gilbert would let her flee, knowing that she would have to do it if they wanted to keep this pretense of friendship free from his growing feelings.  It certainly wouldn’t be the first time it had happened.

But for now, this was okay. Matthew was going to be okay, too. They repeated it in their heads, a simultaneous and silent mantra.

When Marilla called Gilbert’s phone later that evening, Ann had already been pulled down by the last purples of the sunset into sleep. She didn’t stir when the folksy melodies had turned to the tritone chime of his ringtone. Gilbert, confident that Ann was deep in the reprieve of a dream, answered the call.

“Hello?” There was a pause, then - “Oh, hey Miss Cuthbert. No, no, everything’s okay. We didn’t hear the house phone because we’ve been on the porch...Yeah, she’s asleep.” There was another pause, a sigh of relief from Gilbert that carried an entire day’s weight with it. “That really is great news, Marilla. I’ll tell her as soon as she wakes up. We’ll be here when you get home. Is there anything you needed done before then?...Are you sure?...Yeah, you too. Bye.”

Gilbert set his phone down and pressed a kiss to Ann’s hair.

“Look at that, Ann-girl,” he whispered into perfumed strands. “Looks like Matthew’s going to be okay after all.”

//

Ann believed that if the world was against her, she had acclimated to its cruelty. She had developed a sixth sense for predicting whether a single moment would tear apart the peace of the present, or bring days worth of joy.

When Gilbert’s name lit up across her phone at three in the morning, paired with the chimes of a phone call, Ann’s sixth sense told her to steel herself.

“Hey Gil,” she answered, voice groggy. “Everything alright?”

She was met with silence for a few seconds, long enough that Ann began to wonder if Gilbert had really meant to call her at all. Maybe he’d been dreaming or slept with his phone in his hand and -

“Ann, can you -” his voice broke off and she heard him swallow.   “I’m sorry to wake you up.”

The strain in his voice was enough to stir her awake completely, and she sat straight up in bed.

“Gilbert, what’s wrong?” She heard a sharp inhale, a few indistinct voices in the background, some strange beeping noises, then a shuddering exhale.

“My dad was...he’s….he was in an accident. Th- There was nothing they could do.”

Ann deflated as if a massive weight had fallen on her chest. She pulled the phone away from her face, almost as if to hide the whimper that came from her lips and the tears welling up in her eyes. Gilbert’s father was all the family he had left.  There were no uncles, no grandparents, no long lost cousins.

And now there was just Gilbert - the last, the only. Her heart split down the center at the thought of him living the way she’d had to, orphaned and lonely.

“Gilbert, I…” A tear slid down her cheek and she swallowed the lump in her throat. “Where are you? I’m already on my way.”

She found him in a waiting room of the Carmody hospital, thirty minutes outside of Avonlea by car. He was slumped in a chair against the wall in the corner of the sterile space, pale faced and red eyed. Ann waited in the doorway, wondering if she should break his quiet grieving, only to have him look up through heavy lashes.

Ann didn’t have to be told what to do then. In a moment she was kneeling before him and wrapping him in an embrace that she hoped would shield him from the anguish closing in around him. His stiff arms came around her in an instant, his face pressed into the comfort of the crook of her neck.

“It’s alright, I got you,” she soothed. Gilbert let out a quiet whimper fingers digging into the soft fabric of her shirt.

They stayed like that for a while, Ann rubbing his back and soothing him as he wept. She couldn’t ask him what happened, only able to ask one of the passing nurses once Gilbert had gotten up to use the bathroom and wipe off his face. John Blythe had been in a car crash driving home from his job late that night, the nurse told her.

“There are only two types of people on the road that late,” the nurse said. “Third shifters and drunks.”

Ann rubbed her hands over her face and sighed.

“Ann,” Gilbert called quietly out behind her. “Can you take me home?”

She looked back to the nurse, unsure if there was any paperwork to be filled out or procedures to be completed, but the nurse nodded.

They drove home in silence, Gilbert’s cheek pressed against the window of the car, glassy eyes watching the blurry trees pass them on the highway. Ann kept her fingers on the wheel, trying her best to keep her focus centered on the snowy January roads. The car had grown cold, so Ann reached a hand over to turn up the heat and face the vents toward Gilbert, who’d forgotten to take a coat when he left the house.

“I don’t know how to plan a funeral,” he admitted quietly.

“I do,” Ann said, “I’ll help you. I’ll write the obituary and call the hospital and funeral home in the morning.”

Gilbert nodded his head, then turned to look at her.

“At home. We’ll have the funeral at home.”

“Whatever you want,” Ann said, pulling into his driveway. The gray house was all shadows when the pair walked up the front porch steps, Ann’s hand entwined with Gilbert’s to keep him standing. She released his hand and watched him collapse on the couch, face turned away from her. She stood across the room for a few seconds, watching his chest rise and fall with an odd, unsteady rhythm. But then, as if a switch had been flipped in her mind, she began to work.

Caring for Gilbert was much easier than she could have anticipated, not because his pain was less than she expected, but because her heart knew his needs without having to be told. She knew that he was most comfortable when he had his own pillow and the large blanket his mother quilted for him during her pregnancy. Ann wrapped him in his quilt and placed the pillow beneath his head wordlessly, wiping a stray tear from the corner of his eye as she stood up.

Gilbert watched as she moved quietly around the room and turned down pictures that had his father in them, knowing that seeing them would hurt too much. Even in his grief, or maybe because of it, he had a strange, quiet realization. Ann Shirley-Cuthbert was quite possibly the most beautiful thing he’d ever laid eyes on, with her messy bun of auburn hair and strength in her brave shoulders. Bathed in the moonlight coming in through the windows, he could stared at the milky skin of her neck and the tear trails on her cheeks.

Then she came to his side with a plate of peanut butter toast and a shot of whisky.

“How’d you know where that was?” he murmured in a scratchy voice, nodding down at the shot glass. He brought the copper substance to his lips and let the burn travel down his throat.

“I’ve watched you sneak it out a few times,” she confessed. “I brought you some toast in case you’re hungry.”

He wasn’t, but he took a bite out of it to soften the worry lines on her forehead.

“Thanks,” he said, mouth dry from the peanut butter. “You can go home now, Ann. I’ll be fine.”

“If you think I’m leaving you now, Gil, you’ve got another storm coming,” she said, running her fingers through his hair. “If you want to be alone, I can go up to the guest room.”

“No, I don’t want to be alone.”

Ann nodded, standing up to take the chair next to the couch, but Gilbert opened his arms, causing Ann to pause. With no room on either side of him, she settled with her chest pressed to his, legs tangled together. She thought back to the day of Matthew’s surgery, how she’d wanted Gilbert to hold her just like this in her own sadness. That day she’d been too distracted to notice how her heart raced in his chest. Now she was acutely aware of the effect of his breath in her hair and the intimacy of her heartbeat thumping to the same tempo as his.

She thought he’d fallen asleep when she murmured into his shirt, “Do you think you’ll be okay, Gil?” To her surprise, his embrace tightened and she felt a tired sigh blow through her hair.

“Someday,” was his whispered reply.

//

The wake, funeral, and reception went by in a blur to Ann and Gilbert, who played hosts to dozens of bodies coming and going through the Blythe household. Ann stayed by Gilbert’s side throughout the four days, knowing how exhausting it must have been to spend the last weeks of winter break in mourning. Marilla and Matthew helped too - Marilla bringing by meals and clean clothes for Ann, Matthew coming to fix the wood furnace in Gilbert’s living room when it malfunctioned hours before the reception. Diana came by to help clean the house for a few hours because, You’re supporting Gil, Ann, but who’s supporting you? And when it was all finally over, Ann felt like she could release a breath she had been holding onto since Gilbert called to pick him up from the hospital.

Perhaps she relaxed a little too soon.

“I’m sorry, what?” she choked out. She and Gilbert were sitting on the docks of the Lake of Shining Waters, the pond that separated the Barry and Cuthbert lands. Ann’s face was white, even paler with the sunlight reflecting off of the snow and onto her cheeks.

“It’d just be for a year, Ann. I just have to get out of here for a little bit,” Gilbert said, placing his hand on her shoulder. “Walking through Avonlea is like walking beside the ghosts of my past.”

“I’ve haven’t left your side for the past two weeks and you haven’t thought to tell me about this?”

“I knew if I told you I was thinking about it, you’d react like this.”

“And how am I reacting?”

Gilbert took a deep breath and gave a melancholy smile.

“Heartbroken enough that it’d make me consider staying.”

A small little sob escaped her lips and she stood up. She looked out over the frozen pond with its icy fractals, puffs of hot breath blowing fog in front of her face.

“No, I won’t ask you to stay. I know why you have to leave,” she said finally, wiping her cheeks.

“Just think about the sorts of cool souvenirs I can send back from an internship on a cruiseliner. The first stop is Trinidad, you know.”

She turned back to him, biting her lip to keep from smiling. There was no staying angry at Gilbert Blythe for long.

“You’ll call?” she asked.

“Everyday.”

“And send pictures?”

“As many as you want.”

“And when you come back, you’ll finish school?”

“It’s just a gap year, Ann. I’m not waving the white flag yet.”

Ann crossed her arms over her chest and set her face into his shoulder. She hadn’t expected this turn of events, otherwise she’d have cherished his company more, paid more attention to making lasting memories. He brought a hand up to her head and ran his fingers through her hair in a way that was so very Gilbert.

“I’ll miss you too,” he said gently. “I can’t thank you enough for everything you’ve done since Dad…”

“You’ve done the same for me,” she said, pulling back.

“Still, I appreciate it.”

Their gazes lingered on each other’s for a few seconds, bringing back that same warmth that had started blooming in Ann’s chest whenever she really looked at him. How easy it would be to just rise onto her toes and press her lips against-

“Well, I better start packing,” Gilbert said, clearing his throat. Ann blinked a few times, turning her heated cheeks toward the ground.

“Do you want some help?”

An affectionate spark lit up in his eyes.

“I wouldn’t mind some company. The house is a little lonely.”  

Two days later, a small crowd of Gilbert’s favorite people followed him to the Public Bus station. He carried two suitcases with him, his other belongings already mailed to the cruise liner that was to be his home for a year. Ann walked in pace with him at his side, with Diana and their friend Jeri trailing behind. The rest of their friends would be meeting them there, Charlie and Moody, Ruby and Jane.

“I didn’t think everyone would spend their last day of break saying goodbye to me,” Gilbert admitted as he laid eyes on the crowd waiting for him.

“Everyone loves you, Gilbert. Some more than others,” Jeri said, pushing a long strand of brown hair out of her face. She gave Ann a sneaky, sly grin, only to be shot daggers in return.  Before Ann could say anything terribly embarrassing, the group at the bus station exploded with They’re here! and There you are! You finally quit dragging your asses!

Ann was quiet as everyone said their goodbyes to Gilbert, who was nearly rendered speechless at the overwhelming explosion of affection on his behalf. His eyes lingered over to her every few seconds, noticing her unusual silence as easily as if she’d been yelling. When she was the only person left to say goodbye, he walked up to her and stuffed his hands in his pockets.

“Whatcha thinking about, Ann-girl?”

“I’m wondering who I’m going to study with, or who’s going to come annoy me at Patty’s Place when you’re gone.”

“I’m sure Roy Gardner will be happy to fill my shoes.”

“Big pass,” she groaned, nudging him in the stomach with her elbow. “It’s gonna be a long year without you, Gil.”

“You too,” he replied in a reverent murmur. “But I’ll call and text and send pigeon mail and smoke messages just as promised.” She opened her mouth to say something, then closed it, but he could tell there was something waiting on the tip of her tongue. “What is it?”

“I’m about to do something I probably shouldn’t do in front of all of our friends,” she stated seriously.

Gilbert felt his heart leap into his throat, and he swallowed it back with difficulty. There was nowhere else to look but down at Ann, red hair in two braids down the front of her sky blue winter jacket and freckles like snowflakes floating on her cheeks. The chatter or their friends had either fallen silent or he’d simply grown unable to hear it. Ann rose her brows as if asking for permission, and he nodded, entranced and grinning.

Then she was holding the sides of his face and kissing him. Tension drained from them the second their lips made contact, like a long carried weight finally gone from their shoulders. Gasps came from their friends, but neither minded, content to stay held in a tender embrace.

When she pulled back, Gilbert had to remind himself where he was, what he was doing. The look in her eyes was almost enough for him to reconsider the whole internship and stay home to kiss her as long as he wanted.

“They just gave last call for boarding,” she reminded him quietly. Gilbert nodded, not really hearing her. Ann laughed. “That means get on the bus, you idiot!”

Gilbert blinked, looking around at their smug friends.

“ Oh! Right. I’m going. See you guys soon.” He picked up his bag, turned around, took two steps, then turned right back around. Ann, who had deflated the second he’d gone, let out a small gasp when he marched right back up to her.

“Let me take you to dinner when I get back,” he said bravely. Ann let out a half hysterical laugh and covered her face in her hands.

“Okay, okay! Just get on the fucking bus. It’s going to leave without you.”

“You will?” he asked, crooked grin on his face.

“I said I would!” she laughed, then horror crossed her face. “Gilbert, they’re closing the back doors, get on there!”  

He pressed his lips to her cheek, gave one last wave to his friends, and jogged onto the bus. It pulled off before they could find the window he sat by and give their last goodbyes. Then it had pulled out of the station, a distant speck in the horizon, Diana pulled an arm around Ann’s shoulders. She might’ve said something, but Jeri cut her off.

“Well, it’s about damn time,” she stated.

“Leave her alone, Jer,” Diana scolded, then tugged Ann closer to her side.“Come on, babe, let’s get you home.”

//

The kitchen smelled of dried cranberry and crush rose petals several mornings later, the sun dripping in through the translucent cream curtains. When Ann came down the creaky stairs, she found Marilla working over the stove with her back turned to her. The older woman had her long gray hair tied in a single braid down her back, a style which made people who didn’t know her assume she was some sort of nonconformist.

Ann stood in the doorway, enjoying the swell in her heart at the comforting sight of Marilla at work. Then she pulled her phone out and snapped a picture of the scene for days when it was raining and lonely in Kingsport.

“Morning, Ma,” Ann said lowly, as not to startle her. The nickname was one that had originated from Ann’s pressing desire to address Marilla as Mom, and Marilla’s insistence that Ann merely call her by the name her parents had given her.

“Good morning, Ann,” Marilla replied, wiping her arm against her sweaty brow. “You’re just in time. Can you hand me the rose oils? I can’t read the small print on the bottles.”

Ann swept across the fragrant kitchen over to the counter, where Marilla had her open case of essential oils.

“Who are you making soap for this time?” she asked, rifling through the tiny vials in search for the rose colored one.

“I’m making a large batch this time. I’ll be donating some to the church for their craft sale, but you may take the extras and send them to Gilbert if you’d like.”

“Ah, found it!” Ann said triumphantly, handing Marilla the oils. “Is that your long winded way of suggesting that Gilbert isn’t bathing?” Marilla sent a glare over her shoulder, mixing the soap in the warm pan. “No, Gilbert doesn’t like soaps that are too sweet. Diana might like some, though! She’s been having a hard time at home.”

Marilla turned off her mixer.

“Why’s that? Her parents aren’t fighting, are they?”

“No, it’s not that. They just...share different opinions with her on certain things, I guess. It breaks her heart to see her parents talk the way they do.”

“You don’t ever feel that way about me, do you?” Marilla asked carefully. Ann draped an arm around Marilla’s back and leaned her head on her shoulder. She took a deep breath, inhaling the aromatic perfume of the soap, then handed Marilla the bowl of dried berries and petals.

“Not even a little.”

At that moment, the back door swung open with a creak, followed by familiar heavy footsteps. Matthew appeared then, wiping his hands on his jeans and smiling at his girls.

“I see Rachel convinced you to make that soap, after all.”

“She reminded me of my ‘Presbyterian duty’ and was more than happy to remind me of all the filthy people just waiting to be cleaned by the soap of the Lord.”

“Oh I see,” Ann said with a nudge. “You just wanted her to shut up.”

Marilla chuckled, turning off the heat on the stove.

“You watch how you talk about her today. She’ll be here any minute now and you know how that woman doesn’t knock before making herself at home.”

“Wait, why is Mrs. Lynde coming over?” Ann grabbed a piece of Wonderbread from its bag and stuffed it into her mouth. “Did someone die recently? Get pregnant? Find their long lost twin on Eharmony and have tear jerking reunion?”

“Rachel doesn’t always come over to gossip, Ann,” Marilla scolded.

“Come on, Marilla, you have to admit that she’s only ever over when she wants to talk shit about people.”

“Language, Ann,” Matthew said with a cup of morning coffee at his lips. Ann knew he wasn’t terribly upset, since he hadn’t even bothered to look up from his newspaper.

“If you must know, Miss Shirley Cuthbert, Rachel is coming over to drive me to the optometrist. I have an eye surgery today. I won’t be able to drive afterwards.”

“You didn’t tell me you were having surgery done,” Ann murmured. “Is it serious? Why can’t Matthew drive you?”

“It’s nothing to worry about. Matthew has things to accomplish and Rachel needs to get out of the house every now and again,” Marilla insisted, pulling off her crafting apron and folding up. “Now, don’t you have somewhere to be soon?”

She gestured down at Ann’s outfit, a tie dye shirt and a pair of boyfriend jeans with a tiny flag sticking out of the back pocket. The flag and shirt featured three colors - fuschia, purple, and blue. To top it off, she had a pin above her heart that read Kiss Me, I’m Bi!

“Oh Marilla, my first pride officially out of the closet!” Ann said excitedly. “I just wish everyone could be so lucky.”

“That’s why you’re going today, Ann-girl,” Matthew said, sticking his mug in the sink and then pressing a kiss into her red hair. “Lots of people don’t know that’s okay to be who they are, but you’re an expert in that.”

“I suppose I am,” she agreed quietly.

“Stay safe. Call me if you want to come home and I’ll pick you up,” Matthew said. Ann was about to utter her thanks when Rachel Lynde came bursting into through the door. She pulled off her flashy sunglasses, sticking them in her purse, then took one look at the college student standing unashamedly in the kitchen. Then she turned right to Marilla.

“The kids of today are losing their minds,” Rachel remarked.

“Oh thanks Mrs. Lynde,” Ann said sarcastically, “I’m only standing right here. ”

“I’m just saying that-”

“Rachel, do I need to I remind you about Harmon Andrews’ party back in ‘73 when you and Nancy McLean -”

“ Marilla! Rachel choked out, but the damage was done. Ann’s brows were raised into her hairline as she tried, and failed, to take the image of Rachel’s sapphic experiences out of her mind. “We’ll be late for your appointment!”

Rachel grabbed Marilla by her wrist, dragging her toward the back door.

“I guess we’re off. Have a nice time, Ann!” Marilla called, grabbing her purse from the back table before she could be completely kidnapped.

“I suppose that means I should get going, too,” Ann said to a red faced Matthew. “I’ll be back to make dinner.”

  She was standing on the park staircase handing out various flags to empty handed passersby when she saw him. He was a lanky fellow, long limbs and honey colored hair. He had some sort of book in his hands, bounded in a mustard yellow fabric, and judging from the fluid motions of his pencil, he was sketching. Ann only noticed him because every few seconds, he’d peer up at her, then snap his eyes back down to his paper before she could think twice about it. Sitting a few steps down, he kept his bag above him to rest his elbow on. The messenger bag had a few tiny buttons on it, one of which was a thumbnail sized rainbow pin.

Ann left her station for a short moment, and took a few steps down to crouch by him. His eyes went wide when he noticed her before him, and watched nervously as she pulled a rainbow flag from her hands and handed it to him.

“Happy pride,” she said warmly.

“Thanks,” he murmured back, pulling his book against his chest.

“Sorry, I wasn’t trying to look at what you were drawing,” Ann said. “You just looked so lonely over here all by yourself.”

The boy averted his eyes to the pavement but gave a genuine smile that sent a familiar wave of warmth through Ann - the kind that accompanied an interaction with any new kindred spirit. He tugged his book from his chest and handed it to her.

“I hope you don’t mind.”

Ann’s jaw dropped when she saw just what the boy had been drawing. He’d been drawing her - every freckle, every smile line, every loose strand of hair. In fact, Ann didn’t think all the instagram staging or filters in the world could ever make her look so beautiful. There was something raw and ethereal about his sketch, something that made Ann want to be the girl on the paper.

“I...I don’t know what to say. It’s amazing.”

“Sorry I drew you without your permission,” he said meekly. Ann shook her head.

“There’s nothing to apologize for. I’m honored you chose me as your subject. I’m sure there are prettier girls to draw-”

“Aw, come on, that’s not true,” he cut in. Ann shrugged.

“At any rate, thank you for showing me.” Her smile lingered on a moment as she gave him a second to either continue the conversation or return to his work. Bright blue eyes blinked at her, but the boy said nothing. “I should let you get back to it.”

Just as she turned to leave, she heard, “I’m Cole.”

Ann felt a relieved laugh escape her lips; today was not to be a day of lost kindred spirits, after all!

“I’m Ann,” she introduced, sticking a freckled hand in his face, “No E. Though if it were up to me, I’d spell it with an E. Fits my aesthetic a bit more, ya know?”

“I think I can understand that,” he said, smiling as Ann settled down beside him.

“Here, take a handful,” she said, pulling some more flags out of her pocket. “That is, if you want to help me hand them out?”

“Yeah, sure!” The more he spoke, the more be blossomed into happiness, like a flower that needs like but has been kept under shadows too long. “I tried to sign up to work the event, but I couldn’t sneak out of the house without my mom knowing. Even today, she thinks I’m on a field trip for school.”

Ann gave a sputtering laugh.

“It’s the middle of June!”

Cole shrugged. “My mother isn’t known for being the brightest crayon in the box.”

“Speaking of which, you’re an artist?”

A red hot warmth covered Cole’s face, as if he were ashamed to admit it. He pulled the sketchbook back out and opened it to the first page.

“Kinda I guess. I’m not as good as some people.”

Star-struck at the beautiful works in his soft journal, Ann flipped through the pages with gentle fingers and a tender eye.

“I don’t think you’re giving yourself enough credit. These are spectacular. You’re an artist for sure.”

“Are you an artist too?”

Ann shrugged, thought about it, then shook her head.

“My medium is language, words strung together into pretty constellations of poetry and stories,” she said with a flair.

“So...a creative writer?”

“Ding ding ding! But I’m not good enough to make anything of it. Now you, on the other hand, I think you’re good enough to do whatever you want.”

“Maybe someday,” he said, knowing he had to take some of the compliment or risk disappointing her. “Umm, actually, there is- well, that is, if you’re interested. You can say no! I realize that maybe you wouldn’t want to-”

“Cole!” Ann laughed. “Out with it!”

“I’ve been trying to find someone to model for me so that I can practice more portraits. I think I could really make some decent commission money doing them, but I haven’t drawn many - er, women.”

Ann’s face had fallen with shock, and for a second, Cole thought he’d offended her.

“You know what, that’s okay, I shouldn’t have asked,” he murmured quick under his breath. He handed her the flags and moved to pack his things, but she placed a hand on his wrist.

“Wait. I’m not upset you asked me, Cole, I’m just...I was serious when I said there are prettier girls to draw. My looks are nothing special.”

“Well I beg to differ. I look at pretty things for a living and before we met, I was looking at you. Maybe it can help us both out. I can show you how you look through the eyes of other people.”

“I know already how people look at me.”

“I mean the ones that count.”

Ann flipped back to the sketch Cole had been drawing of her minutes ago, and stared at it for a second. Then she made the mistake of looking up at his hopeful eyes, the ones that longed for a kindred spirit for too long, the ones that had taken this one risk.

“Fine, I’ll do it. But it’s your fault for choosing me if your pieces come out looking odd.” Cole only smiled.

"I'll risk it."

//

They met every Tuesday in the late afternoon. Ann chose the time because she said it was when the sun was directly outside her window, bathing her the pastel turquoise of her room with “the most beautiful golden light in all of Avonlea.” She could have chosen three in the morning seven days a week for all he cared, he just wanted out of his house. Besides, if he’d brought a girl home, he knew for sure that his mother would say, “Cole MacKenzie, did you finally get over that homosexual phase you were in?”

Ann’s home was one were he felt safe, the first few visits showing him all he needed to see of Marilla and Matthew Cuthbert, the kind siblings who’d adopted Ann.

“How long ago did you come out to Marilla and Matthew?” Cole asked one day, not taking his eyes off the intricate details of his sketch. Ann herself sat in the window seat, cross legged and looking off to the right.

“Matthew knew from the first day,” she confessed. “I was sitting outside my group home in Bolingbroke. One of the boys had just teased me about having a crush on one of the girls from school. Really, I think he was just projecting a bunch of his bullshit onto me. Matthew originally intended to adopt him - he and Marilla had planned on adopting a boy, and James was the only guy in the house - but then he met me and the plans changed. Matthew heard all about my crush at our first meeting, among other things. I talked his ear off.”

“And it didn’t bother him even a little?” Cole couldn’t imagine the unconditional acceptance of a parent, not with the way things were in his household.

“Nope. He never dated women, so I’ve often wondered...But in the end, his acceptance of an LGBT youth was what convinced my case worker to assist him with the adoption. The rest is history. I told Marilla several months later, and was officially out in Avonlea earlier this year. All of it made for a very undramatic coming out story.”

“You’re one of the lucky ones,” he murmured bitterly.

“I am. I’m blessed,” she replied sincerely. “But Cole, you’re my friend now. You’re part of this family, whether you like it or not. You’ll be one of the lucky ones soon.”

Cole smiled at this, considering offhandedly that she was right about the sunset in her window. In that moment, she looked like a fiery angel, fierce and strong.

“I’m already one of the lucky ones,” he decided.

The moment was broken by Ann’s phone chiming with a recognizable little chime. Ann didn’t budge, but only moved her eyes to see her phone sitting on the seat beside her. A smile erupted on her face and she broke her pose, swiped across her screen, and held the phone up.

“Hey stranger!” she said.

“ Hey yourself, carrots.” He was as lovely as she remembered, with those soft brown curls and warm hazel eyes. His face lit up as soon as he’d seen her, and Ann was sure she mirrored the expression. It’d been so long since he’d had time to call, giving her plenty of time to remember parting at the bus station.

“You’re lucky I’ve missed you too much to acknowledge that abominable you just called me,” she said sweetly. “Oh, Gil, how are you?”

Instead of Gilbert answering, she heard another voice come in from the background.

“ Alright Blythe, the shower is yours, but don’t take too lon-” A face appeared in the screen, bearded and dark eyed. “ Is that your girl there? Ann?”

Gilbert swatted the man away as Ann laughed, “Guilty!”

“ Can’t a guy make a call home in peace? Ann, that’s Bash, one of the guys I met working here.”

Cole came around and poked his mop of blonde strands into the frame.

“Any friend of yours is a friend of mine,” Ann said. “This is Cole, the friend I told you about from pride.”

“Oh hey, man! Nice to finally put a face to a name! ”

“Cole, this is Gilbert, my…” Ann gaped for a second, causing Gilbert to raise his brows. “This is Gilbert.”

“I’ve heard lots of good things about you,” Cole said with a smile. “Medical man, right?”

“ The very same,” Gilbert replied.

“Hey, Gil, I thought you were rooming with that Nova Scotia man. What was his name? Matthew? Marcus?”

“ Maddox,” Gilbert offered. “ Bash’s roommate was being a racist asshole, and so the room director let us switch.”

“ The man didn’t want to shower in the same place I had. Can you imagine?” Bash cut in.

“Oh, I think I could,” Cole grumbled the same time Bash called out, “Oh, tell her the news, man!”

“News?” Ann said carefully. “Everything alright?”

“ No no, everything’s great. Seriously, Ann, you’d be the first to know if something was wrong. I have a feeling you’d feel a disturbance in the bosom connection between the two of us.”

“Now wait a second-”

“ But I called to let you know that my supervising doctor onboard is thrilled with my performance the last few months. He wants to get in touch with a colleague at the University of Toronto. You know, set me up an interview so that I could meet the board and get a head start on planning for grad school. It’s an amazing opportunity, and a great connection to have. Plus, the University of Toronto is one of my top choices.”

“That’s great, Gil, but isn’t that...you know, really far away?”

“Not any farther than Trinidad and the rest of the Caribbean.”

Ann bit her lip and forced herself to smile. While Gilbert was off saving lives and delivering babies, where would she be? In the back of his mind?

“Don’t forget about the small people when you’re becoming a big fancy doctor.”

“Forget about you, Queen Ann? Never.” Ann blushed, feeling the same way that she might if he suddenly told her he was in love with her - heart racing, stomach fluttering. Suddenly the image on the screen shifted away to a very passionate face of a very passionate Bash.

“ Oh Ann, I wish I could tell you of my plans for you once I graduate medical school! I’m going to be a biiiiig fancy doctor and we’ll get a biiiiig fancy house.”

“Hey!” Gilbert cried. The image on the screen turned into a rollercoaster as Bash swung the phone away from Gilbert’s grabby hands.

“ And we’ll get married and have teeny, weeny little spitfire babies. Twenty of them!”

“ Sebastian! I’m serious!”

“No? How’s twenty-five?”

Ann exchanged an awkward look with Cole, who’s smirk gave off tangible energy.

“Gilbert’s cute,” he murmured knowingly.

“ See!” Bash laughed.

Finally Gilbert was able to snag the phone away from his obnoxious roommate, and his distressed face greeted Ann when he finally managed to steady his hand.

“ Sorry about that. ”

“Not at all,” Ann said, shaking her head. “It’s just nice to hear from you, even in embarrassing circumstances.”

“I know I haven’t called much lately. I’ll fix that.”

“Effective immediately?”

“Yes ma’am,” Gilbert said officially with a solemn nod of his head. “Listen, I have to get back to my post soon. I really will call. Next time I want to hear all about how Diana’s doing and  Marilla’s eye surgery, okay?”

“Yeah, okay,” Ann said, forcing her voice to stay even. “Hey, Gil, you know…” If it was Marilla, Matthew, or Diana, the call would have ended with Ann’s typical I love you. But she couldn’t say that now, not to Gilbert. Not because it wasn’t true, in fact, each day she knew more and more that it was true. The fact that it was true made it terrifying, especially now that he was thinking of going to Toronto. Cole grabbed her hand where Gilbert couldn’t see it, seeing some of her thoughts across her face.

“ What is it? ” Gilbert probed.

“Take care of yourself, yeah?” she said finally.

“Love you too, Shirley.”  Ann’s heart gave a pleasant little jump. If only he were home. “Talk to you later.”

She smiled right as the phone beeped and went black. Ann heaved a heavy sigh and threw her phone onto her bed.

“Well, I feel like my life is complete now that I’ve finally met Gilbert Blythe: the man, the myth, the legend,” Cole said dramatically.

“Oh please, it’s just Gilbert,” Ann said, settling back into her pose. Taking the cue, Cole grabbed his sketchbook again and sat in front of her.

“But he’s not just Gilbert to you.”

Ann sighed and gave Cole a surrendering look.

“No, no he’s not.”

[joroanblog]

Essentials and also current advances of two dimensional- polyacrylamide gel electrophoresis

Abstract

  Gel- based proteomics is one of one of the most flexible methods for fractionating protein facilities. Amongst these approaches, 2 dimensional- polyacrylamide gel electrophoresis (2-DE) stands for a pillar orthogonal strategy, which is widely used to all at once fractionate, determine, and measure proteins when combined with mass spectrometric identification or various other immunological examinations. Although 2-DE was first introduced greater than 3 decades earlier, a number of challenges and restrictions to its utility still exist. This testimonial goes over the concepts of 2-DE in addition to both recent methodological advancements as well as new applications. Introduction

  Two dimensional polyacrylamide gel electrophoresis (2-DE) is taken into consideration a powerful device made use of for separation as well as fractionation of complex protein combinations from cells, cells, or other organic samples. It enables splitting up of hundreds to hundreds of proteins in one gel. This strategy became extra popular and also detailed after the prime advances and high resolution 2-DE adjustment [1] In fact, the modification established by O'Farrell enabled 2-DE method to deal with up to 5000 protein- standing for areas in an even 2 dimensional circulation and allowed exact splitting up of protein places with high precision [2] 2-DE technique is considered to be among the leading powers in the growth of proteomics as well as protein studies. It provides the initial step for further evaluation of the differentially controlled protein spots using mass spectrometry and western blotting. Making use of 2-DE has actually been efficiently defined in a lot of cases to reveal both physiological systems as well as healthy proteins associated with professional pathologies that can aid in the exploration of biomarkers. Basic principles of 2-DE Idea of 2-DE

  2-DE is composed mainly of two actions of separation; first measurement and also 2nd dimension. In the very first measurement, protein molecules are resolved relying on their isoelectric factor (pI) [2] Separation of proteins under a pH slope enables intense band recovering using different techniques such as immobilized slope electrophoresis (IPEG), isoelectric focusing (IEF), or non-equilibrium pH slope electrophoresis (NEPHGE). In the 2nd measurement, protein splitting up is done based on molecular weight making use of SDS Laemmli or Tris-Tricine barriers. Due to the fact that it is unlikely that different protein particles may have the very same physicochemical buildings (pI and also MW), proteins are successfully separated by 2-DE rather than 1D-SDS PAGE [2] A prominent advantage of 2-DE is that the resolution gotten throughout the very first dimensional separation is not missed in the second electrophoresis when IEF gel strip is linked to the SDS- PAGE gel [2, 3] Applications and energies of 2-DE

  2-DE is an effective and widely utilized technique for evaluation of intricate protein mixes with exceptional ability to separate hundreds of proteins at once. It provides direct aesthetic verification of changes in protein/post-translational modifications (PTMs) wealth, thus offering early reason for downstream logical actions with discovering post- and also co-translational alterations, which can not be forecasted from the genomic series. Other applications of 2-DE consist of whole proteome analysis [4], cell differentiation [5], discovery of biomarkers as well as illness pens, drug discovery, cancer research [6], bacterial pathogenesis [7], purity checks, microscale protein purification, as well as product characterization. Advantages and toughness of 2-DE Effectiveness

  Throughout the last few years, a number of methodological enhancements have added to enhance the toughness of 2-DE workflows. Using incapacitated isoelectric concentrating (IEF) strips, ampholytes- based buffers, very delicate dyes, as well as gel imaging software application made the irregularity most likely from upstream process such as protein loss throughout extraction [8] In a recent multi-laboratory research on the expediency of 2-DE, it is reported that 70-93% of areas were discovered with coefficient of variant (CVs) less than 20% within same laboratory scientists [9] On the other hand, 72% of areas revealed CVs with less than 20% throughout labs [9] This searching for verifies the feasibility and also the effectiveness of 2-DE. In addition, 2-DE becomes much less variable when multiplexing electrophoresis developed. Differentially identified examples perform at the exact same time decreased the opportunity of artefacts arised from technical errors. Lastly, the recent improvements of the gel image analysis lessened the former high portion of place identification failure estimated to get to 60% which thought about among the significant factors to irregularity seen with 2-DE [10] Visualized mapping analysis

  One of the one-of-a-kind features of 2-DE is its ability to deal with undamaged full-length proteins (approximately 5000 protein) in a solitary gel. This includes pictured detection of the physico-chemical homes such as MW and pI with possible metrology based on the spot intensity [8] Healthy proteins of interest could be characterized using peptide mass finger printing or when penetrated with antibodies. Moreover, 2-DE enables separation as well as identification of PTMs and also protein isoforms (Figure 1). In several instances, PTMs could be identified by horizontal or vertical moving of a protein place as these adjustments usually change the protein MW and also pI [11] Compatible platform for further evaluation

  2-DE gel easily and also successfully couples with numerous various other evaluation and also biochemical techniques. Hence, it supplies a suitable platform for succeeding evaluation. For example, stained gels can be followed by place excision, destaining, protein extraction, food digestion, and also evaluation of the tryptic peptides by mass spectrometry. Although coomassie blue could be reversibly destained as well as suitable with mass spectrometry, silver discoloration, it is not suitable as a result of the usage of formaldehyde or glutaraldehyde during the repairing and also sensitization step that causes lysine deposit cross-linking within the protein chain interfering MS analysis and thus will certainly prevent trypsin digestion [12, 13] Numerous alterations in the silver nitrate stain technique were performed to overcome this downside. Compatibility of 2-DE consists of bottom-up proteomics to determine proteins and identify their amino acid sequences or additionally continued by the top-down proteomics (shotgun) in which the crude healthy proteins remove is absorbed directly for evaluation. In an additional powerful mix, antibody- based evaluation could be coupled before or after 2-DE For example, immune-affinity filtration can be utilized to pre-fractionate a protein of rate of interest before running 2-DE such as phosphorylated [2] or ribonucleoproteins [14] Many commonly, 2-DE fractionated proteins are subjected to either in-gel food digestion to prepare tryptic peptides for mass spectrometric analysis or gels are confirmed for protein of passion using western blotting [15] Limitations of 2-DE. Low reproducibility

  In the traditional method of the initial measurement IEF, the service provider ampholytes is used to build pH slope. The provider ampholytes-based pH slope made from soft unsupported tube gels (commonly 4% acrylamide) is not stable. There is set to batch irregularity and vulnerable to cathodic drift (a progressive loss of fundamental proteins throughout lengthy running of electro-focusing under electric area), causing low reproducibility as well as calling for mindful surveillance of electric field [2] Replacement of carrier ampholyte-based pH gradient in tube gel with the incapacitated pH slopes (IPG) was the essential advancement in boosting the reproducibility of 2-DE [16, 17] Difficulty in separating hydrophobic and also extremely acidic or fundamental proteins Various sorts of healthy proteins can constantly be missing out on, because of the trouble in separating membrane- bound (hydrophobic) and severe healthy proteins [18, 19] Especially, extremely acidic or basic proteins are neither conveniently removed neither solubilized. This problem in removal depends primarily on the solubilization power of the buffer utilized in the IEF step. Numerous initiatives have actually been carried out for better solubilization of membrane layer healthy proteins using different chaotropes or cleaning agents. For immediate, Triton X- 114 and also CHAPS showed a powerful recuperating power in the direction of hydrophobic proteins on 2-DE when checked immunologically [20] An additional study reported the use of cationic detergent benzyldimethyl-n-hexadecylammonium chloride to boost settling hydrophobic healthy proteins with GRAVY index as reduced as 0.56 [21] Comparable renovation was reported when using detergents such as DHPC [22] and also 1,4-dithioethanol [23] Although solubilization of wide variety of healthy proteins could be achieved using denaturating option (Urea and also Thiourea) and zwitterionic detergents (SB 3-- 10) [24], resistance of specific healthy proteins still stays an integrated problem [2, 24] Slim vibrant variety of 2-DE

  Reduced vibrant series of healthy proteins is one of the tough problems run into during 2-DE. Very plentiful peptides mask reduced plentiful ones, which may be reflective of reduced plentiful healthy proteins. In addition, the visualization of pale protein areas (reduced bountiful) separated on 2-DE gels is likewise governed by tarnishing sensitivity. For example, classic coomassie has a narrow vibrant array with discovery limitation of only regarding 100 ng. Colloidal coomassie is fairly higher in level of sensitivity with discovery limit 10 ng. The vibrant array could be enhanced to a discovery degree below 1 ng with the availability of highly sensitive silver-staining method [12] and a variety of fluorescence dyes such as SYPRO-Ruby and Deep purple flurophore dyes (Figure 2). Consequently, making use of delicate tarnish boosts protein sample dynamic range, causes effective gel imaging, and also finally leads to successful mass spectrometric recognition and immunological recognition [11] Additionally, exhaustion of highly abundant proteins such as albumin and also hemoglobin considerably improves the dynamic range of 2-DE by permitting better focusing and also mass spectrometric picking (see Prefractionation, enrichment, and deficiency before 2-DE section). Therefore, the limited detection sensitivity of 2-DE does not handle the real dynamic series of protein concentration in cell and tissue removes, or biological liquids [2] Various other service to get rid of the dynamic array issue is to fill more protein example and also using a huge 2-DE gel (24 centimeters). Nonetheless, this method could lead to manufacturing of jammed photos with non well-separated spots. Reduced throughput and also labor- intensiveness

  2-DE is labor-intensive and also has a loved one reduced throughput. The throughput of 2-DE is adequate for lots of small standard research study studies, however it may provide a major challenge for jobs that involve testing of a great deal of scientific examples. In addition, 2-DE requires skills and experience to combat any feasible problem capturing because of the procedural artifacts. Much care needs to be thought about to decrease the variations throughout example processing that lasts up to 3 days in larger gels. Recent advancements and also modern technologies related to 2-DE 2D- DIGE

  The growth of image technology has actually presented differential imaging gel electrophoresis (DIGE) strategy. This method was designed in an effort to boost sensitivity and reproducibility of 2-DE making use of multiplexed fluorescent dyes- labeled protein samples. 2D-DIGE is based mainly on running more than one example (optimum 3 samples) on a single gel simultaneously to attend to the concern of gel-to gel irregularity. In this technique, different fluorescent cyanine (Cy) dyes are used for labeling proteins from various examples [25] After blending these examples in equal proportion and also running them together as one sample, very same protein from different samples migrates to the exact same position on the 2D gel where it could be conveniently explored and differentiated by the different fluorophore-labeled color and also imaged to determine its wealth. 2D-DIGE is an important device, specifically for medical research laboratories involved in the resolution of protein expression levels and disease biomarker discovery. When outright biological variant between samples is the main objective, as in biomarker discovery, 2D-DIGE is just one of the approaches of option. Prefractionation, enrichment, and also depletion before 2-DE

  Protein sample prefractionation before 2-DE has been executed to reduce sample complexity. Because of this, the reduced wealth healthy proteins existing in these fractions will be clearly stood for in 2-DE. Proteins identified in the prefractionated examples have a higher number of peptides. Additionally, low molecular weight healthy proteins can be clearly detected when example intricacy is minimized [26] Prefractionation eventually increases filling capability of samples onto the 2-DE gel and also results in much better resolution, visualization and also recognition. Test prefractionation can be done in several various methods, broadly subdivided right into three levels: mobile, subcellular and also protein subfractionation [27] For example, mobile extraction is a generalized method to screen the entire proteome of an offered example. A removal barrier having urea and also thiourea with NP-40 generally recuperates most healthy proteins [24] In subcellular prefractionation, a thickness gradient (isopycnic) centrifugation technique [28], or additionally tissue filters as well as buffers technique is utilized to homogenate a given tissue followed by using careful solvents to dissolve mitochondrial, ER-golgi, or nuclear proteins [29] Protein subfractionation based upon its physico-chemical residential properties is an additional degree of fractionation. This method could be made use of to fractionate proteins based on their charge such as strong cation exchange (SCX) approach [30] or based upon their pI such as Liquid stage IEF Zoom ® prefractionator [31] Enrichment of protein of rate of interest that exists in the sample in low wealth such as phosphorylated healthy proteins (represents 3-4%) might be crucial before 2-DE. For example, in one of our experiments to identify tyrosine phosphorylated glomerular healthy proteins related to slit diaphragm, samples were immunoprecipitated making use of protein A- Sepharose then subjected to 2-DE [32] Relying on the nature of the example, biological liquids such as plasma as well as urine may require elimination of specific proteins (albumin or hemoglobin) to increase the resolution of splitting up before 2-DE. On the downstream mass spectrometric degree, the excess of highly tryptic peptides created from these abundant proteins bias the identification in the direction of these healthy proteins on the cost of lower bountiful ones. In contrast, depletion increases peptides record opportunities of reduced bountiful healthy proteins [33, 34] Blue native Gel electrophoresis for membrane layer proteins studies

  Blue-Native polyacrylamide gel electrophoresis (Blue Native PAGE) was initially presented by Schagger and von Jagow as a method for separating enzymatically energetic membrane protein complexes under mild condition [35] In this approach, the anionic dye Coomassie Brilliant Blue G-250 (5% w/v) is mixed with the protein sample prior to gel loading. This color has the capacity to offer negative charges to the surface area of the protein. It induces a fee change that boosts solubilization of hydrophobic proteins especially membrane intrinsic electron/proton transfer facilities in mitochondria giving an international analysis of membrane proteomics. The dye binds to protein complexes as well as both move during electrophoresis. Lastly, the gels are stained with coomassie dye once more prior to mass evaluation. A current application for this strategy was productive in identifying specific substances within protein intricacies of inflammasomes [36] as well as integrin and also histone facilities in placenta [37] In addition to its capability to unveil protein intricacies, this technique permits much better splitting up of hydrophobic healthy proteins (membrane proteins) also [38] 2-DE for article translational modifications (PTMs).

  The evaluation of protein post-translational adjustments (PTMs) has actually become an important topic for the research study of cell biology, condition treatment, as well as disease avoidance. 2-DE supplies a straight monitoring of protein PTMs in gels in addition to its relative abundance. Publicly readily available online devices such as ProMoST [39] and also JVirGel [40] could be used for that function. In addition, specific staining dyes for PTM surveillance were established and also ended up being extensively applied in the proteome researches. In phosphoproteome study, molecular Probes, Pro-Q ® Diamond phosphoprotein gel tarnish is a development innovation that gives a precise technique for uniquely tarnishing phosphoproteins in polyacrylamide gels. Likewise, for glycoproteomics, readily available fluorescent dye; Lissamine rhodamine B sulfonyl hydrazine (LRSH) was presented to particularly stain the glycoproteins. This tarnish relays on periodate/Schiff base mechanism. Unlike traditional approaches used for the characterization of PTMs such as the enrichment methods either with antibodies or paralyzed material, lectin's binding strategy, or chemical based techniques, 2-DE deals a logical device with high resolution and also high reproducibility by taking the benefit of the well known adjustment in pI and also MW caused by several modifications [41] Outlines on the kinds and also current schedule of 2-DE.

  First dimension electrophoresis. The initial dimension electrophoresis can be performed making use of ionic substances, which reacts as acid or base and also is termed as provider ampholyte pH slope. It could be blended and enhanced for vast or more limited pH arrays. Numerous techniques for IEF could be applied in the first measurement electrophoresis as discussed below. Conventional IEF.

  The traditional approach of IEF depends upon the carrier ampholyte where proteins migrate in an option media up until getting to the stability state when its web charge equals to no. Proteins that are in a pH area below its isoelectric factor (pI) will be positively billed and will move in the direction of the cathode. As it migrates with a gradient of boosting pH, nonetheless, the protein's total charge will reduce until the protein reaches the pH region that corresponds to its pI. Now, the migration stops. As a result, the proteins end up being concentrated into sharp fixed bands with each protein positioned at a point in the pH slope corresponding to its pI. [2] Although this traditional technique is simple to prepare and also do not require much casting tools, it has a major drawback as the ampholytes have some susceptibility to stream towards cathode as well as this slope flow typically creates a decrease in the reproducibility. Immobilized pH slope (IPG).

  Paralyzed pH gradient strip (IPG) is an integrated part of polyacrylamide gel matrix fixed on a plastic strip. Co-polymerization of a set of non-amphoteric buffers with different chemical properties is included [2, 42] A prepared- made IPG strips are available with different lengths and also pI. Usually, short length IPG strips are used for quick screening while much longer one for maximal and thorough analysis. Various designs of 2- DE gels are received Figure 3. An industrial pre-casted acrylamide gel matrix co-polymerized with a pH gradient on a plastic strip causes a secure pH worth over the conventional ampholyte approach. It has an ability to stay clear of cationic build-up and also to generate a better-focused protein with much less smearing [2, 42] There are lots of various other advantages of using IPG strips over ampholytes such as decreased cathodic drift, greater mechanical toughness as the strips are casted on a plastic support that reducing gel damage, as well as greater protein packing ability because of the example loading technique [17, 43] look at this site electrophoresis protein

Business Name: Helena Laboratories Address: 1530 Lindbergh Dr, Beaumont, TX 77707, United States Phone No: +1 409-842-3714

[thebibliomancer]

Essential Avengers: Avengers #118: To The Death

December, 1973

Dormammu strikes me as a guy that would be more interested in ‘to the pain.’ And I’m not one to criticize cover combat strategies but maybe don’t all jump at Dormammu in one easily Evil Eye’d clump.

Last time: Dormammu tricked the Defenders into collecting the Evil Eye, Loki tricked the Avengers into ineffectually trying to stop them. The two teams finally realized something was up but not in time to stop Dormammu from stealing all the Evil Eye bits and merging Earth with the Dark Dimension.

This time: Things are bad forever. Buildings are warping, people are turning into monsters and not in the cool way.

But no matter the odds, the Avengers and Defenders will fight

to the death, yes.

God, Namor. I don’t know what you were going for but it probably wasn’t constipation. And got to love Cap trying for an intimidating flex.

And then we zoom out a little and the assembled heroes look less mighty and dynamic and more royally screwed.

They’ve got a couple royals among them so yeah, royally screwed sounds right.

Geez. Look at that disproportionally huge upper body monster man and his tiny legs. Look at that girl turning into a dragon feets up. Lookit everything being on fire and buildings sagging like a souffle.

This is some good ‘world gone to hell’ art.

The Avenging Defenders and Defending Avengers get to work holding back the monsters. But they have to hold back because these were people once and will be people again if they can thwart Dormammu.

Dr. Strange casts a quick spell to prevent the two superhero teams from turning into monsters. Sadly, that’s all he can protect without expending too much energy to face Dormammu.

It really kind of drives home the shit that the teams are in. But for a roll of the dice, they could have been the ones turning into monsters.

Now, the only way to stop this is to stop Dormammu. And the only way to stop Dormammu is to go to his dimension. I guess since Earth isn’t fully merged he has to stay on his side until the hour is up.

But Cap raises the point that they can’t just desert Earth. Hundreds of people could be killed in their monster forms before they could fix things. Hell, the monster people might kill people who have yet to transform.

But they can’t split the party. Dormammu plus Evil Eye means they need to confront him with their full strength.

What is a superhero to do? Leave the few to possible death to save the many? There’s no easy answer.

Not unless SHIELD shows up to lend a hand. Hey, just like Age of Ultron!

Now, monsterfying is happening all over the world but SHIELD is an international agency at this point so hey! Problem solved. SHIELD will take over the Los Angeles situation so the Avengfenders can deal with the big picture.

Now, yeah, debate cut short with a convenient answer but I’m glad Englehart addressed that there would be collateral damage. That the heroes couldn’t just pop off and fight a fire-headed jerk without leaving behind a bad situation.

I like it was Defender Dr. Strange who said they needed to go after Dormammu and Avenger Captain America who worried about leaving defenseless civilians behind. That feels about right for where the teams would respectively focus.

Anyway, Dr. Strange casts his dimension jump spell and off the two teams go. While SHIELD begins a holding action. Although, I’m not sure how you perform a holding action when an entire world is going bonkers and your own people might turn into monsters.

Because, yeah. That happens. SHIELD didn’t get a fancy protection spell against monsterfying.

Countess Valentina Allegro de Fontaine transforms into a big lizardy monster and turns on Nick Fury. He stuns her but realizes that anyone else could turn at any time. This holding action is doomed if the heroes don’t come through.

SPEAKING OF, welcome to the Dark Dimension, Avengers and also Defenders. Hope you survive the experience.

And the Dark Dimension is weird and, yes, mostly poorly lit.

Thor wants all the fliers to fly ahead but Dr. Strange cautions him not to stray from the gray, meteor pocked trail. The laws of reality are weird in the Dark Dimension and its not safe off the trail.

Some of the Avengers are resentful to take Dr. Strange’s lead but they all charge forward along the gray path.

Meanwhile, miles or millenia ahead, Dormammu shoots off rainbows from the Evil Eye to amuse himself.

Loki is unimpressed. Because he can’t see anything. So maybe Dormammu should fulfill his part in the bargain and restore Loki’s sight now? After all, Dormammu couldn’t have pulled this off without Loki’s aid.

Dormammu thinks no. In fact, he thinks its kennel time for Loki and puts him in a mystic pink cage. See, once he got the Evil Eye, he realized that Loki had tried to screw him over by alerting the Avengers.

So he can stay in the cage and think about what he’s done.

AND THEN THE WATCHER SHOWS UP! Oh but don’t fret. He’s not here to interfere. He’s just here to watch. Two dimensions merging together? That’s a pretty sweet spectacle.

I guess Dormammu appreciates an audience because he promises enough spectacle to justify the Watcher’s entire existence of watching things.

Also, Dormammu swears by his sister. Is her head also on fire?

Meanwhile, the Avengers and Defenders hit a snag. The path just kind of ends. Also: now they’re being attacked by the Mindless Ones. Good news though: the Mindless Ones brought their own path so there’s the way forward.

Feels kind of like a side-scrolling beat-em-up. You hit the end of the path. A bunch of dudes show up that you have to beat up and then it lets you continue forward.

Problem is that physical combat just makes the Mindless Ones excited. So instead all the Avengers that have ranged blasty powers all line up and blast at the same time, causing the Mindless Ones to flee off-panel.

But they left their sweet pink road.

Mantis, who apparently has an internal clock rivaling the Andalites, tells the group that they only have twenty minutes left. I say rivaling because apparently she ‘literally feels each second being ripped from the cosmos around her by the invisible hands of time!’

That sure is a power to have, Mantis.

Meanwhile, lets check in on the homefront. Buncha heroes fighting monsters and reacting to this bazonkers event. I think the best might be Dracula objecting to Earth becoming monstery because it might affect him. AND THAT MUST NEVER BE!

Or Luke Cage grumbling that this was supposed to be his day off. Later on he would try to take Martin Luther King Day as a personal day only to get wrapped up in Vision and Scarlet Witch’s problems. Poor Luke.

Also Gorgon doesn’t know Triton’s name. What’s funniest about that is that the marvel wiki tries to justify it as in-universe sarcasm. Sometimes people mess up, guys.

Meanwhile, the Dark Dimension. The Avengers and Defenders have finally reached Dread Dormammu.

Unfortunately, he has a shield up to keep the Mindless Ones away from himself. Dr. Strange can sunder the shield but it will take time. And they probably don’t have a lot of that left.

Also, think about the Watcher’s vow of non-intervention. I think its doomed to failure. Because just by showing up, he influences events even a little. People generally know that something is significant when the Watcher shows up. In the Infinity Gauntlet event, Thanos knew that a big battle was headed his way because the Watcher showed up to watch. Granted, Thanos was near omnipotent at the time so its not like it affected things much there.

So lets look at here instead. The Watcher shows up to watch. Thor sees the Watcher and shouts excitedly because its someone he knows because Thor is always jazzed to see a friend. Dormammu suspects that this excitement means the Watcher was lying about being neutral. The Watcher goes ‘how dare, non-intervention is my game, dawg. I love these guys but I’ll stand by and watch them brutally slaughtered, that's how much I’m about non-intervention.’

And then Dormammu tells the Watcher to chill out.

But while this conversation was going on, Dr. Strange bursts through Dormammu’s barrier. You could argue that the distraction the Watcher provides just by being present is an intervention. The Watcher is a bad anthropologist, probably. At the least, he should invest in a cloaking device.

Anyway, Dormammu isn’t worried. Why, with the Evil Eye backing him up he can merely wave his hand and instantly K.O. the Defenders.

Alas.

Still, the Defenders have been trouncing the Avengers this whole crossover. Time for them to pull their own weight. Plus, the final part of this story is taking place in an Avengers book.

The Avengers are uncowed and shout that thing they love to shout and rush forward.

So Dormammu turns the ground into quicksand.

Because if you have power overwhelming, why not mess with your enemies. Its not like they can stop you, probably.

Except for Scarlet Witch, Iron Man, and Thor all the Avengers fall victim to the quicksand. 

Scarlet Witch wants Thor and Iron Man to try to pull the rest of the Avengers free but Swordsman tells her not to stop for them.

He’ll use the heat rays from his sword to fuse some solid islands out of this quicksand to give the bogged down Avengers something to hang onto.

... Okay. Not sure if that’s how quicksand works but whatever.

Iron Man, Thor, and Scarlet Witch run forward to confront Dormamu. Dormammu is okay with this. In fact, he likes the humanity of these superheroes. Makes them more fun to play with. In fact, they’re so human why not become all human.

And he vanishes Iron Man’s armor and turns Thor into Donald Blake.

Kind of weird that Scarlet Witch is still running around. Whats the deal?

She was born with her power. So she can’t be separated from it, at least not in the same way that affected the other two.

Of course, Wanda phrases this differently. And also says “You cannot stop me!” So 10/10 for confidence there.

So instead Dormammu attacks Scarlet Witch with a harmless little rain shower. NOT! Its a glue storm. And now she’s covered in glue.

Dormammu is being weirdly non-violent with his ways of stopping the Avengers.

I guess because its all a game to him. Although I fear its because he doesn’t want to break his new toys just yet.

But with all the Avengers incapacitated in one form or another, there’s nothing to stop Dormammu from winning in abouuuuuuuuuuuut... five minutes. Oh, sorry. Five of your Earth minutes. Dormammu minutes are presumably cooler.

But Loki has been waiting for an opportunity. Dormammu’s attention is fully taken up with gloating over the Avengers and not paying any to Loki in his mystic cage.

So Loki busts out one of his less used powers and transforms into an insect to fly between the bars of the cage. Relying on the sound cues of Dormammu’s ceaseless yammering, Loki grabs Dormammu from behind and tries to pry the Evil Eye away from him.

I wonder if he learned any good wrassling moves from growing up with Thor.

Now Scarlet Witch is the one who has been forgotten. Still pretty ensnared in goo, she takes the opportunity to unleash all of her stored power at the Evil Eye.

Because its thought-controlled. And from what I gather, usually has to be triggered by touch. But with the probability warping power of Scarlet Witch’s arbitrary mutant power, she causes the Evil Eye to trigger itself.

And the Evil Eye eats Dormammu. And, uh, sort of goes off right in Loki’s face.

Due to probabilities gone mad, the energy discharge fulfills Loki’s most fervent hope and restores his sight to him. It just... has a minor side effect of driving him totally insane.

With Dormammu defeated, the dimension merge is cancelled. The people unmonsterfy, the landscapes return to normal, and a confused Ben Grimm takes credit for fixing everything.

Back in the Dark Dimension, the Watcher congratulates the Avengers and Defenders and decides to explain some things.

Apparently Dormammu’s extra-dimensional form is composed of raw energy that is constantly on fire. This energy is generated by belief and worship of Dormammu, like with most mystic entities. So that’s how the Evil Eye ate him. It probably can’t eat anyone not made of energy. Probably.

But Dormammu will be back. And not even in the ‘this is comic books and he’s an iconic villain, of course he’ll be back’ sort of way. More in the “those who love sin will continue to call upon him -- and in time, their reverence will reshape him!”

Way to screw the rest of us over, all you sin lovers.

As for Loki. He had Dormammu’s energy blasted through his brain. And no brain from Earth’s dimension can cope with so much sorcerous force. His mind has broken under the strain, reverting him to the mentality of an infant.

I don’t know if ‘totally insane’ is in fact the right way to describe that?

And since the Watcher has been handing out free exposition like its going out of style, it would be nice if the Vision answered a question. Why did he panic instead of fly when Dormammu made the ground into quicksand?

Vision can’t answer the question because he doesn’t understand it himself. Yet another weird clue in some long mystery about the Vision.

But anyway. The day is saved thanks to the Scarlet Witch mostly. So Dr. Strange scoops up the Evil Eye.Time to go back to Earth and use it to fix Black Knight. Also he just up and erases everyone’s memory of seeing Iron Man and Thor’s civilian identity so they can reveal it when they choose.

WHICH IS NICE I GUESS BUT WOULD HAVE BEEN NICER IF DR. STRANGE HAD ASKED FIRST! He just erases everyone’s minds without a by your leave.

And he half-asses it too because Iron Man and Thor presumably still know each other’s identities since they found out independently.

I get that status quo is god and maybe the decision to have Iron Man and Thor’s identity be revealed should happen in their respective books but instead of having mind erase happen, just make it so nobody saw their faces? The Defenders were unconscious, the Avengers were mostly stuck in the mud. Really only Scarlet Witch would have seen and recognized them and she can be trusted with a secret.

Hell, that would be a good thing to follow up this story with. Thor and Iron Man having a quiet conversation with Scarlet Witch after this to formally entrust her with their secret identity.

Alas.

So that was the Avengers/Defenders War. The Avengers go on from this to have a Halloween adventure at the Rutland Halloween Parade. Remember last time they went to Rutland, straw feminism kicked their butts.

The Defenders go on to try to unstone Black Knight with the Evil Eye to mixed success. Its technically counted as the final part of the Avengers/Defenders War despite the war being over. Maybe I’ll cover it but basically the Defenders end up transported to the 12th century Crusades, discover the Black Knight living through the body of one of his ancestors, and have to stop Mordred from getting the Evil Eye. Prestor John ends up with the Evil Eye, implying a weird time loop. And Black Knight decides to stay in the past times so he can continue fighting in the Crusades and I guess eventually meets Exodus, Mysterious Overpowered X-Men Villain #71.

So. The Avengers/Defenders War. It was more of an Avengers/Defenders Scavenger Hunt. And the Avengers really underperformed until the end when Scarlet Witch grabbed the Quidditch and won the game for Earth.

I mentioned that it reminds me a lot of JLA/Avengers and I’ll go into that a little now. We have two teams clashing where the Avengers are way outmatched. We have them fighting over a number of arbitrary powerful items. We have the conflict being engineered by outside manipulators. We have all the arbitrary powerful items being taken away from the heroes once they’re gathered and used to make the world go screwy. And we have the two separate teams spending some time to get to know each other, decide that they’re not so bad, and then teaming up to kick a cosmic jerk in the face. And also, Hawkeye is a member of both the fighting teams.

These very broad points also correspond to JLA/Avengers.

Its probably a coincidence but I wonder if the Avengers/Defenders War was used as an inspiration for how to handle an extended crossover between two unmatched teams. JLA/Avengers was written by Kurt Busiek and he’s kind of a continuity buff.

I know that the ‘two superhero teams fight because villain pulling the strings’ thing has become a cliche but a hero vs hero story where the teams are able to just hang out and socialize and part on good terms feels good.

[yesdanielblisslove]

Essentials and also current advances of two dimensional- polyacrylamide gel electrophoresis

Abstract

  Gel- based proteomics is one of one of the most flexible methods for fractionating protein facilities. Amongst these approaches, 2 dimensional- polyacrylamide gel electrophoresis (2-DE) stands for a pillar orthogonal strategy, which is widely used to all at once fractionate, determine, and measure proteins when combined with mass spectrometric identification or various other immunological examinations. Although 2-DE was first introduced greater than 3 decades earlier, a number of challenges and restrictions to its utility still exist. This testimonial goes over the concepts of 2-DE in addition to both recent methodological advancements as well as new applications. Introduction

  Two dimensional polyacrylamide gel electrophoresis (2-DE) is taken into consideration a powerful device made use of for separation as well as fractionation of complex protein combinations from cells, cells, or other organic samples. It enables splitting up of hundreds to hundreds of proteins in one gel. This strategy became extra popular and also detailed after the prime advances and high resolution 2-DE adjustment [1] In fact, the modification established by O'Farrell enabled 2-DE method to deal with up to 5000 protein- standing for areas in an even 2 dimensional circulation and allowed exact splitting up of protein places with high precision [2] 2-DE technique is considered to be among the leading powers in the growth of proteomics as well as protein studies. It provides the initial step for further evaluation of the differentially controlled protein spots using mass spectrometry and western blotting. Making use of 2-DE has actually been efficiently defined in a lot of cases to reveal both physiological systems as well as healthy proteins associated with professional pathologies that can aid in the exploration of biomarkers. Basic principles of 2-DE Idea of 2-DE

  2-DE is composed mainly of two actions of separation; first measurement and also 2nd dimension. In the very first measurement, protein molecules are resolved relying on their isoelectric factor (pI) [2] Separation of proteins under a pH slope enables intense band recovering using different techniques such as immobilized slope electrophoresis (IPEG), isoelectric focusing (IEF), or non-equilibrium pH slope electrophoresis (NEPHGE). In the 2nd measurement, protein splitting up is done based on molecular weight making use of SDS Laemmli or Tris-Tricine barriers. Due to the fact that it is unlikely that different protein particles may have the very same physicochemical buildings (pI and also MW), proteins are successfully separated by 2-DE rather than 1D-SDS PAGE [2] A prominent advantage of 2-DE is that the resolution gotten throughout the very first dimensional separation is not missed in the second electrophoresis when IEF gel strip is linked to the SDS- PAGE gel [2, 3] Applications and energies of 2-DE

  2-DE is an effective and widely utilized technique for evaluation of intricate protein mixes with exceptional ability to separate hundreds of proteins at once. It provides direct aesthetic verification of changes in protein/post-translational modifications (PTMs) wealth, thus offering early reason for downstream logical actions with discovering post- and also co-translational alterations, which can not be forecasted from the genomic series. Other applications of 2-DE consist of whole proteome analysis [4], cell differentiation [5], discovery of biomarkers as well as illness pens, drug discovery, cancer research [6], bacterial pathogenesis [7], purity checks, microscale protein purification, as well as product characterization. Advantages and toughness of 2-DE Effectiveness

  Throughout the last few years, a number of methodological enhancements have added to enhance the toughness of 2-DE workflows. Using incapacitated isoelectric concentrating (IEF) strips, ampholytes- based buffers, very delicate dyes, as well as gel imaging software application made the irregularity most likely from upstream process such as protein loss throughout extraction [8] In a recent multi-laboratory research on the expediency of 2-DE, it is reported that 70-93% of areas were discovered with coefficient of variant (CVs) less than 20% within same laboratory scientists [9] On the other hand, 72% of areas revealed CVs with less than 20% throughout labs [9] This searching for verifies the feasibility and also the effectiveness of 2-DE. In addition, 2-DE becomes much less variable when multiplexing electrophoresis developed. Differentially identified examples perform at the exact same time decreased the opportunity of artefacts arised from technical errors. Lastly, the recent improvements of the gel image analysis lessened the former high portion of place identification failure estimated to get to 60% which thought about among the significant factors to irregularity seen with 2-DE [10] Visualized mapping analysis

  One of the one-of-a-kind features of 2-DE is its ability to deal with undamaged full-length proteins (approximately 5000 protein) in a solitary gel. This includes pictured detection of the physico-chemical homes such as MW and pI with possible metrology based on the spot intensity [8] Healthy proteins of interest could be characterized using peptide mass finger printing or when penetrated with antibodies. Moreover, 2-DE enables separation as well as identification of PTMs and also protein isoforms (Figure 1). In several instances, PTMs could be identified by horizontal or vertical moving of a protein place as these adjustments usually change the protein MW and also pI [11] Compatible platform for further evaluation

  2-DE gel easily and also successfully couples with numerous various other evaluation and also biochemical techniques. Hence, it supplies a suitable platform for succeeding evaluation. For example, stained gels can be followed by place excision, destaining, protein extraction, food digestion, and also evaluation of the tryptic peptides by mass spectrometry. Although coomassie blue could be reversibly destained as well as suitable with mass spectrometry, silver discoloration, it is not suitable as a result of the usage of formaldehyde or glutaraldehyde during the repairing and also sensitization step that causes lysine deposit cross-linking within the protein chain interfering MS analysis and thus will certainly prevent trypsin digestion [12, 13] Numerous alterations in the silver nitrate stain technique were performed to overcome this downside. Compatibility of 2-DE consists of bottom-up proteomics to determine proteins and identify their amino acid sequences or additionally continued by the top-down proteomics (shotgun) in which the crude healthy proteins remove is absorbed directly for evaluation. In an additional powerful mix, antibody- based evaluation could be coupled before or after 2-DE For example, immune-affinity filtration can be utilized to pre-fractionate a protein of rate of interest before running 2-DE such as phosphorylated [2] or ribonucleoproteins [14] Many commonly, 2-DE fractionated proteins are subjected to either in-gel food digestion to prepare tryptic peptides for mass spectrometric analysis or gels are confirmed for protein of passion using western blotting [15] Limitations of 2-DE. Low reproducibility

  In the traditional method of the initial measurement IEF, the service provider ampholytes is used to build pH slope. The provider ampholytes-based pH slope made from soft unsupported tube gels (commonly 4% acrylamide) is not stable. There is set to batch irregularity and vulnerable to cathodic drift (a progressive loss of fundamental proteins throughout lengthy running of electro-focusing under electric area), causing low reproducibility as well as calling for mindful surveillance of electric field [2] Replacement of carrier ampholyte-based pH gradient in tube gel with the incapacitated pH slopes (IPG) was the essential advancement in boosting the reproducibility of 2-DE [16, 17] Difficulty in separating hydrophobic and also extremely acidic or fundamental proteins

  Various sorts of healthy proteins can constantly be missing out on, because of the trouble in separating membrane- bound (hydrophobic) and severe healthy proteins [18, 19] Especially, extremely acidic or basic proteins are neither conveniently removed neither solubilized. This problem in removal depends primarily on the solubilization power of the buffer utilized in the IEF step. Numerous initiatives have actually been carried out for better solubilization of membrane layer healthy proteins using different chaotropes or cleaning agents. For immediate, Triton X- 114 and also CHAPS showed a powerful recuperating power in the direction of hydrophobic proteins on 2-DE when checked immunologically [20] An additional study reported the use of cationic detergent benzyldimethyl-n-hexadecylammonium chloride to boost settling hydrophobic healthy proteins with GRAVY index as reduced as 0.56 [21] Comparable renovation was reported when using detergents such as DHPC [22] and also 1,4-dithioethanol [23] Although solubilization of wide variety of healthy proteins could be achieved using denaturating option (Urea and also Thiourea) and zwitterionic detergents (SB 3-- 10) [24], resistance of specific healthy proteins still stays an integrated problem [2, 24] Slim vibrant variety of 2-DE

  Reduced vibrant series of healthy proteins is one of the tough problems run into during 2-DE. Very plentiful peptides mask reduced plentiful ones, which may be reflective of reduced plentiful healthy proteins. In addition, the visualization of pale protein areas (reduced bountiful) separated on 2-DE gels is likewise governed by tarnishing sensitivity. For example, classic coomassie has a narrow vibrant array with discovery limitation of only regarding 100 ng. Colloidal coomassie is fairly higher in level of sensitivity with discovery limit 10 ng. The vibrant array could be enhanced to a discovery degree below 1 ng with the availability of highly sensitive silver-staining method [12] and a variety of fluorescence dyes such as SYPRO-Ruby and Deep purple flurophore dyes (Figure 2). Consequently, making use of delicate tarnish boosts protein sample dynamic range, causes effective gel imaging, and also finally leads to successful mass spectrometric recognition and immunological recognition [11] Additionally, exhaustion of highly abundant proteins such as albumin and also hemoglobin considerably improves the dynamic range of 2-DE by permitting better focusing and also mass spectrometric picking (see Prefractionation, enrichment, and deficiency before 2-DE section). Therefore, the limited detection sensitivity of 2-DE does not handle the real dynamic series of protein concentration in cell and tissue removes, or biological liquids [2] Various other service to get rid of the dynamic array issue is to fill more protein example and also using a huge 2-DE gel (24 centimeters). Nonetheless, this method could lead to manufacturing of jammed photos with non well-separated spots. Reduced throughput and also labor- intensiveness

  2-DE is labor-intensive and also has a loved one reduced throughput. The throughput of 2-DE is adequate for lots of small standard research study studies, however it may provide a major challenge for jobs that involve testing of a great deal of scientific examples. In addition, 2-DE requires skills and experience to combat any feasible problem capturing because of the procedural artifacts. Much care needs to be thought about to decrease the variations throughout example processing that lasts up to 3 days in larger gels. Recent advancements and also modern technologies related to 2-DE 2D- DIGE

  The growth of image technology has actually presented differential imaging gel electrophoresis (DIGE) strategy. This method was designed in an effort to boost sensitivity and reproducibility of 2-DE making use of multiplexed fluorescent dyes- labeled protein samples. 2D-DIGE is based mainly on running more than one example (optimum 3 samples) on a single gel simultaneously to attend to the concern of gel-to gel irregularity. In this technique, different fluorescent cyanine (Cy) dyes are used for labeling proteins from various examples [25] After blending these examples in equal proportion and also running them together as one sample, very same protein from different samples migrates to the exact same position on the 2D gel where it could be conveniently explored and differentiated by the different fluorophore-labeled color and also imaged to determine its wealth. 2D-DIGE is an important device, specifically for medical research laboratories involved in the resolution of protein expression levels and disease biomarker discovery. When outright biological variant between samples is the main objective, as in biomarker discovery, 2D-DIGE is just one of the approaches of option. Prefractionation, enrichment, and also depletion before 2-DE

  Protein sample prefractionation before 2-DE has been executed to reduce sample complexity. Because of this, the reduced wealth healthy proteins existing in these fractions will be clearly stood for in 2-DE. Proteins identified in the prefractionated examples have a higher number of peptides. Additionally, low molecular weight healthy proteins can be clearly detected when example intricacy is minimized [26] Prefractionation eventually increases filling capability of samples onto the 2-DE gel and also results in much better resolution, visualization and also recognition. Test prefractionation can be done in several various methods, broadly subdivided right into three levels: mobile, subcellular and also protein subfractionation [27] For example, mobile extraction is a generalized method to screen the entire proteome of an offered example. A removal barrier having urea and also thiourea with NP-40 generally recuperates most healthy proteins [24] In subcellular prefractionation, a thickness gradient (isopycnic) centrifugation technique [28], or additionally tissue filters as well as buffers technique is utilized to homogenate a given tissue followed by using careful solvents to dissolve mitochondrial, ER-golgi, or nuclear proteins [29] Protein subfractionation based upon its physico-chemical residential properties is an additional degree of fractionation. This method could be made use of to fractionate proteins based on their charge such as strong cation exchange (SCX) approach [30] or based upon their pI such as Liquid stage IEF Zoom ® prefractionator [31] Enrichment of protein of rate of interest that exists in the sample in low wealth such as phosphorylated healthy proteins (represents 3-4%) might be crucial before 2-DE. For example, in one of our experiments to identify tyrosine phosphorylated glomerular healthy proteins related to slit diaphragm, samples were immunoprecipitated making use of protein A- Sepharose then subjected to 2-DE [32] Relying on the nature of the example, biological liquids such as plasma as well as urine may require elimination of specific proteins (albumin or hemoglobin) to increase the resolution of splitting up before 2-DE. On the downstream mass spectrometric degree, the excess of highly tryptic peptides created from these abundant proteins bias the identification in the direction of these healthy proteins on the cost of lower bountiful ones. In contrast, depletion increases peptides record opportunities of reduced bountiful healthy proteins [33, 34] Blue native Gel electrophoresis for membrane layer proteins studies

  Blue-Native polyacrylamide gel electrophoresis (Blue Native PAGE) was initially presented by Schagger and von Jagow as a method for separating enzymatically energetic membrane protein complexes under mild condition [35] In this approach, the anionic dye Coomassie Brilliant Blue G-250 (5% w/v) is mixed with the protein sample prior to gel loading. This color has the capacity to offer negative charges to the surface area of the protein. It induces a fee change that boosts solubilization of hydrophobic proteins especially membrane intrinsic electron/proton transfer facilities in mitochondria giving an international analysis of membrane proteomics. The dye binds to protein complexes as well as both move during electrophoresis. Lastly, the gels are stained with coomassie dye once more prior to mass evaluation. A current application for this strategy was productive in identifying specific substances within protein intricacies of inflammasomes [36] as well as integrin and also histone facilities in placenta [37] In addition to its capability to unveil protein intricacies, this technique permits much better splitting up of hydrophobic healthy proteins (membrane proteins) also [38] 2-DE for article translational modifications (PTMs).

  The evaluation of protein post-translational adjustments (PTMs) has actually become an important topic for the research study of cell biology, condition treatment, as well as disease avoidance. 2-DE supplies a straight monitoring of protein PTMs in gels in addition to its relative abundance. Publicly readily available online devices such as ProMoST [39] and also JVirGel [40] could be used for that function. In addition, specific staining dyes for PTM surveillance were established and also ended up being extensively applied in the proteome researches. In phosphoproteome study, molecular Probes, Pro-Q ® Diamond phosphoprotein gel tarnish is a development innovation that gives a precise technique for uniquely tarnishing phosphoproteins in polyacrylamide gels. Likewise, for glycoproteomics, readily available fluorescent dye; Lissamine rhodamine B sulfonyl hydrazine (LRSH) was presented to particularly stain the glycoproteins. This tarnish relays on periodate/Schiff base mechanism. Unlike traditional approaches used for the characterization of PTMs such as the enrichment methods either with antibodies or paralyzed material, lectin's binding strategy, or chemical based techniques, 2-DE deals a logical device with high resolution and also high reproducibility by taking the benefit of the well known adjustment in pI and also MW caused by several modifications [41] Outlines on the kinds and also current schedule of 2-DE.

  First dimension electrophoresis. The initial dimension electrophoresis can be performed making use of ionic substances, which reacts as acid or base and also is termed as provider ampholyte pH slope. It could be blended and enhanced for vast or more limited pH arrays. Numerous techniques for IEF could be applied in the first measurement electrophoresis as discussed below. Conventional IEF.

  The traditional approach of IEF depends upon the carrier ampholyte where proteins migrate in an option media up until getting to the stability state when its web charge equals to no. Proteins that are in a pH area below its isoelectric factor (pI) will be positively billed and will move in the direction of the cathode. As it migrates with a gradient of boosting pH, nonetheless, the protein's total charge will reduce until the protein reaches the pH region that corresponds to its pI. Now, the migration stops. As a result, the proteins end up being concentrated into sharp fixed bands with each protein positioned at a point in the pH slope corresponding to its pI. [2] Although this traditional technique is simple to prepare and also do not require much casting tools, it has a major drawback as the ampholytes have some susceptibility to stream towards cathode as well as this slope flow typically creates a decrease in the reproducibility. Immobilized pH slope (IPG).

  Paralyzed pH gradient strip (IPG) is an integrated part of polyacrylamide gel matrix fixed on a plastic strip. Co-polymerization of a set of non-amphoteric buffers with different chemical properties is included [2, 42] A prepared- made IPG strips are available with different lengths and also pI. Usually, short length IPG strips are used for quick screening while much longer one for maximal and thorough analysis. Various designs of 2- DE gels are received Figure 3. An industrial pre-casted acrylamide gel matrix co-polymerized with a pH gradient on a plastic strip causes a secure pH worth over the conventional ampholyte approach. It has an ability to stay clear of cationic build-up and also to generate a better-focused protein with much less smearing [2, 42] There are lots of various other advantages of using IPG strips over ampholytes such as decreased cathodic drift, greater mechanical toughness as the strips are casted on a plastic support that reducing gel damage, as well as greater protein packing ability because of the example loading technique [17, 43]   original site serum protein electrophoresis test 

Business Name: Helena Laboratories Address: 1530 Lindbergh Dr, Beaumont, TX 77707, United States Phone No: +1 409-842-3714

[ronaldbosieyiii]

DIY Insulated Dog House

 Things I Used in This Project: ISOtunes Bluetooth Hearing Protection Titebond III Wood Glue Miter Saw Stand Plan Miter Saw Infinity Table Saw Blade Infinity Miter Saw Blade Woodpeckers Speed Square Armor Tool Pocket Hole Jig Woodpeckers Straight Edge Woodpeckers 12″ Square Triton Track Saw Brad Nailer Titebond Thick and Quick

When my dogs aren’t with me in the shop, they are hanging out on the wrap around porch which has plenty of shade, is out of the rain, and always has a great breeze. During the summer it’s awesome because they stay cool but during the winter it’s hard for them to get warm. So this week I built them a dog house with the purpose of giving all three of them a place to get away from the wind and snuggle up. Let me show you how I did it. 

This will be an insulted dog house so instead of going with my first instinct which was a plywood body, I went with more of a framing design made from 2x4s ripped in half. I first sent the 2x4s through my table saw to take off the rounded edges on both sides, then readjusted the fence and cut them directly in half. 

After getting them cut to width I then cut everything to length at the miter saw. 

I started by constructing what will be the two sides, coming up a few inches on a few pieces to give the house some small feet and get it up off the ground. Again, mine will be under a porch and should be kept dry but just incase. 

I’ll be using pocket holes to join things together and I’m very excited to introduce a new jig to hit the market that has some really great features.It’s the Armol Toos Pocket Hole Jig. Right off the bat they made the deck of the jig the same height as a 2×4 so it’s easy to create side support feeds. Of course I don’t need them in this instant but this will come in handy. So on a traditional pocket hole jig you have three things to adjust, the drill block, the clamping tension, and the drill bit collar….on this jig all of those things are self adjusting so all you have to do is stick in your material. If you want to go from 1 1/2” to 3/4” you just clamp the new material into place and you see everything automatically adjusts for you. You can use the onboard Allen key to tighten down on the drill bit collar then you’re ready to go. If you want to double check, there are indication markings on the side annnnd they are color coordinated to the length of screw you need for that thickness of material. Pretty sweet if you ask me. 

After getting all the pocket holes drilled, I started attaching things. Even though this house shouldn’t ever see moisture it will be outside so I went ahead and used Titebond III wood glue since it’s a waterproof glue.

After getting one side assembled, I repeated the process to create another. Next I started working on the front panel, first cutting everything to size, drilling in pocket holes, then gluing and screwing it in place. This design is very easy to adjust depending on your size of dog. Since I have three and they all like to snuggle up together, I made this one long enough to fit three of their beds but still give them room to get around one another. 

After getting the door in place, I thought I should double check to see if they could comfortably fit. With that test passed, I moved on to attaching the back panel. Moving things to the floor to have more working room. 

Next up was a floor, but before putting in the decking material I threw in some insulation. This comes in a big 4×8 sheet and is 3/4” thick. Since my framing is 1 1/2” thick I cut two panels per opening and doubled filled up the cavity. I would use a straight edge and a box blade or my pocket knife to cut it out, then stuck it in place. 

Once the bottom was full I measured and cut some plywood to deck it. I ended up cutting this piece in half to make getting it into place a little easier and I just made sure to cut it in the center so I could join the seam to the floor joist. 

And now more insulation for the walls! To make this step easier I would cheat and set the panel directly on top of the foam then just trace the shape I was needing. If you’re working alone, you can set the insulation on the floor so you have less travel distance to move the entire house. I would line one of the straight edges up to the edge of the insulation then trace the rest, cut it then stick it in place. I was originally thinking I would need to glue these panels in place but the friction fit was so snug I skipped the glue all together. Oh and just a reminder this insulation has foil backing on both sides so if you want to use your table saw to cut it down, and have a SawStop then just be sure to put it in bypass mode or it will tip right away. 

The set in place and tracing method worked so well I next repeated it for the siding! For the siding I’m going with beadboard. This stuff is a huge pain to paint but it does give it some good texture and it’s pretty lightweight, only being 3/8” thick. As you can see I dragged the full sheet over to my work area, then set flipped around the house in order to trace all the sides. If you do this of course just make sure a horizontal reference is squared up to the headboard so your panel doesn’t come out wonky looking. 

After getting the sides traced I used my tracksaw to cut all the shapes out then Titebond III and brad nails to stick it in place. I started by attaching the two side panels so that when I cut and attached the front and back, the edges of the side panels would be covered up.

On the front panel, I went ahead and covered up the opening to the dog house with the bead board, but then after getting it secured in place I used a large drill bit to punch a hole through. Then I used a router and a flush trim bit to perfectly cut out the opening. If you don’t have a router then you can also use a jigsaw or even a reciprocating saw to do this. 

Now moving on to the roof. Since the dog opening is muuuuch smaller than me, I wanted to make the roof into an access panel to the inside so I decided to make it hinged. This will allow me to easily grab the beds to wash, check for snacks, or anything else. I decided to use the left over beadboard I had from the siding to create this roof and also decided to split it into two doors instead of just a single. This is fine for mine since it will live under a covered porch but if you plan to place your house in the rain then keep it as a single panel so rain won’t drip through the seam. 

Before attaching things, I set the roof panels aside and gave everything a coat of paint. Like I said before, beadboard is not fun to paint because it takes two forms of application. I would first use a roller to get all the flat surfaces, but then used a brush to get into all the valleys. Looking back on it, I should have used my sprayer, that would have saved me a ton of time, but oh well. For the body I am going with a grey and while it was drying, I also grabbed what will be the trim boards and threw a coat of paint onto them as well to be drying. Instead of white, I’m going with a light grey. 

While those were drying, I started attaching the roof panels. I started by centering and squaring up the panels to the body. Once I had it just so, I used a pencil to trace the underside of the panels. This line indicates the outside of the body, but since I want to also insulate the roof I needed to know where the inside of the body fell on these panels. With the framing being 1 1/2” thick, I grab something that was 2” thick, which just happened to be a level, and used this to mark the inside line. Now that I had this reference I could measure the area inside and cut a piece of insulation to size. To attach these panels Im’ using a multi surface glue made by Titebond called Thick and Quick. Since I could use brads to hold it in place while that dried, I grabbed an assortment of tools and just weighted it down for a bit. 

While that was setting up to dry I started working on the trim. I ripped down my 1x boards I had painted earlier then started cutting them to length and attaching them. Glue doesn’t stick very well to painted surfaces so learn from my mistake and leave the top edge of your body unpainted to give you a clean surface to glue to. 

I’m again starting on the edges of the house so that the front and back trim will cover up the end grain of the side trim. I also always cheat on trim. Instead of measuring and cutting I typically just hold the board in place and mark the back side. 

By this time the insulation was well past stuck in place so I first attached a portion of paino hinge to the underside then crawled inside the house and attached it to the body. And I should have seen it coming but to be honest I didn’t think about it when I was working the design….but the door actually has a built in stop with the overhang on the back end. So that’s convient. : ) 

Nice, that works nicely. After getting one secure, I repeated on the second. I used the Tounge and groove feature of the beadboard to make the seam in the middle disappear. This is great except when I tried to lift up one without the other, it was running into each other and prevented it. To fix this issue I grabbed my multi tool and just notched out a small portion in the back so that I could open one and give it clearance to pass up the other. There we go, now it’s working correctly. 

Next I did all the finish painting to the inside, the underside of the roof, and any other exposed wood. Now when I was trimming around the door I left the top piece of trim long so I could hang something cute and special. I used my CNC machine to cut out a cut little paw print and instead of just glueing this to the side, I instead added a small hook to the overhung trim and the paw so that I could hang it and there would be a slight swing to it. Hehe, I think the dogs will appreciate the marking. 

I thought I was done but the roof was really bothering me with how thin it looked compared to everything else. So I ended up ripping some 1/2” plywood and adding a small strip around the primereter of the underside of the roof. This just bulks up the look some and I think makes it looks better. 

And that’s it! Now the pups have a place they can crawl inside to get out of the wind that’s always present on our hill. They can all be together which will make them happy, but still have room to get around one another. Also, I have a way to peak in at them to make sure they aren’t staying up late and ordering pizza. 

So a few followers on my Instagram page were mentioning their dogs would tear up the exposed insulation on the inside. Mine don’t chew or scratch things up, but if yours do then I would certainly recommend adding a layer of thin sheeting to the inside as well. Don’t forget I have a set of plans for this build in the plans section of this website.

That’s it for this one, I hope you enjoyed it.

The post DIY Insulated Dog House appeared first on Wilker Do's.

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Astronomers think they’ve found a moon the size of Neptune in a distant star system

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Astronomers think they’ve found a moon the size of Neptune in a distant star system

Nearly eight thousand light-years away from Earth, there’s a star about the same size as our sun. Like our own solar system, that distant star is orbited by a planet about the same size as Jupiter. But that’s where the similarities end. Around that planet circles a Neptune-sized gas giant, which may be the first moon discovered outside the solar system, and the largest moon ever observed.

Over the past 20 or so years, scientists have confirmed the existence of nearly 3,800 exoplanets, or planets around other stars. However, although nearly 200 moons are known to orbit planets in our solar system—Jupiter alone has at least 79—up to now researchers had not yet detected any moons around exoplanets. Their discovery was published today in Science Advances.

To look for such “exomoons,” astronomers from Columbia University examined data from NASA’s Kepler space telescope on 284 transiting planets—worlds that passed between their stars and the observatory, resulting in a brief dimming of the light of those stars. They focused on worlds that took more than 30 days to complete orbits around their stars—prior work suggested that worlds that orbited in less time were likely too close to their stars for any moons to survive.

The scientists detected anomalies hinting at a moon around the exoplanet Kepler-1625b. This gas giant is about the same diameter as Jupiter, and orbits the solar-mass star Kepler-1625 about 7,825 light-years from Earth in the constellation Cygnus the swan.

The researchers then requested and received about 40 hours of time on NASA’s Hubble Space Telescope to analyze Kepler-1625b during its 19-hour-long transit across the face of its star. Using Hubble, which is about four times more precise than Kepler, they detected two sets of telltale signs that suggested the presence of an exomoon. “We indeed conclude that a moon is an excellent explanation for the data in hand,” said study senior author David Kipping, an astrophysicist at Columbia University. (Read more about Kipping’s hunt for exomoons here.)

First, after the exoplanet passed in front of its star, the researchers detected a second and much smaller decrease in the star’s brightness 3.5 hours later. This supports a scenario where a moon trailed the planet like a dog following its owner.

Second, the astronomers found the planet began its transit nearly 80 minutes earlier than predicted. This is consistent with a picture where a moon’s gravitational tug would cause its planet to wobble from its predicted location. Although the gravitational pull of another planet could in principle also cause this anomaly, Kepler found no evidence for additional planets around this star during its four-year mission.

“It sounds like they struck gold,” said astrophysicist Sean Raymond at the University of Bordeaux in France, who did not take part in this work. “Moons are out there and findable. It’s an exciting next step in exoplanet exploration.”

The researchers estimated the exomoon, dubbed Kepler-1625b-i, is only 1.5 percent the mass of its companion planet, a ratio nearly that of Earth and its moon. However, that still means the exomoon may be gargantuan—Kepler-1625b is likely several times Jupiter’s mass, so its moon is likely about the mass and diameter of Neptune.

The size of the moon is surprising—”all the large moons in the solar system are at most about 1/10,000th as massive as their host planets,” Raymond said. The pair are so large that it “could also be regarded as a binary planet system,” said Avi Loeb, chair of Harvard University’s astronomy department, who did not participate in this study.

“The biggest things will be the easiest to find,” Kipping said. “This may not represent a particularly common type of moon system—it’s just it’s for the easiest for us to find.”

The researchers estimated the exomoon orbits about 33 million kilometers from its world. This means it may lie close enough for its planet’s gravitational pull to rip it apart. But there’s nothing like this in our solar system for the researchers to observe directly. To study the dynamics of the planet-moon pair, the researchers created computer models to see how the two might interact. “In about three-quarters of the simulations that we did, we find the moon is perfectly stable,” Kipping said. “We don’t have any reason to believe that orbit is unstable.”

Kepler-1625b orbits its star about the same distance that Earth does the sun. That puts the exoplanet and its moon within their star’s habitable zone—the area around the star warm enough for standing bodies of liquid water to survive. There is life virtually wherever there is water on Earth, so the hunt for alien life often focuses on habitable zones.

Both Kepler-1625b and its potential newfound moon are gas giants, and so cannot support the bodies of water needed for life as we know it to survive. However, “if there are additional rocky moons orbiting the large planet, they might be habitable,” Loeb said. “This is the most exciting prospect for future follow-ups on this discovery.”

The size of this potential exomoon raises questions about how it formed. Some moons, like Earth’s, are thought to have coalesced from the debris of a giant impact against their companion planet. Others, like Neptune’s moon Triton, likely started off as independent bodies only to later get ensnared by their planet’s gravitational pull. However, neither of these scenarios seems to fit this exomoon, said study lead author Alex Teachey, an astrophysicist at Columbia University—it is difficult to see how an impact against a gas giant would split off a moon, or how a moon of such size could get captured.

Another possibility is that this exomoon may have formed from the same material as its parent planet, as is suspected with Jupiter and its moons. New models of moon formation have suggested “a small fraction of moons can end up being as massive as about 1 percent of their host planet mass, which is about the case for Kepler-1625b,��� Raymond said.

In the future, hunts for exomoons will likely look at exoplanets farther away from their stars than Earth is from the sun, Kipping said. These searches with Hubble or NASA’s upcoming James Webb Space Telescope may even turn up exomoons smaller than Jupiter’s largest ones, he added.

The scientists noted they are still urging caution about their find. “The first exomoon is obviously an extraordinary claim, and it requires extraordinary evidence,” Teachey said. “Furthermore, the size we’ve calculated for this moon, about the size of Neptune, has hardly been anticipated, and so that too is reason to be careful here.”

The researchers hope to use Hubble again to monitor Kepler-1625b and confirm the exomoon’s existence.

Written By Charles Q. Choi