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askinkiskarma · 2 years ago

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Illicit Affairs | Chapter X: Invisible String

Pairing: Neteyam x Human/Avatar!Reader

Chapter I Chapter II Chapter III Chapter IV Chapter V Chapter VI Chapter VII Chapter VIII Chapter IX

Synopsis: Back from the dead, you have to heal in order to be able to move on and be forever part of the people. You and Neteyam finally find each other, giving in to desires you've both had for years.

Warnings: SMUT (18+ minors DNI!!!), some angst, fluff, all the feels, so many feels, the end

Word Count: 14,7k words (i need to be stopped)

A/N: The last chapter of Ilicit Affairs is finally here!! It took me a lot longer than I thought to finish this, but that's because this chapter is longer than my first like 5 chapters combined. I don't know what happened hahahah. I'm so excited to share this with you, the conclusion and culmination of this whole story. I hope you find it a satisfying pay-off, and I'd love to hear allll of your thoughts. Honestly, replies and asks make my life and I cry a little bit every time someone interacts with my content. I hope you have enjoyed this journey, and make sure you read til the end for a (hopefully pleasant) surprise ;)

As always, thank you to everyone who asked to be tagged, I love you all sm!! Ok enjoy byeee x

“Isn't it just so pretty to think all along there was some invisible string tying you to me? A string that pulled me out of all the wrong arms right into that dive bar Something wrapped all of my past mistakes in barbed wire Chains around my demons, wool to brave the seasons One single thread of gold tied me to you”

Neteyam stood in the room that he knew so well now, he had memorised its cracks in the white walls, and he was holding your hand, slowly tracing the now-fading yellow bruises along your arm. He looked at the tube that was going into the nook of your elbow and followed his eyes along it, until it reached a little clear pouch with a liquid that was dripping slowly all the way into your body. You were talking animatedly with Kiri, Norm and Max, looking over a bunch of plants displayed on a table in front you. It was incredible to see you, you looked like a completely different person than you were a few days ago. As soon as you woke up, you asked for the treatment you worked on and got working, and your symptoms improved massively. You were still sick, and the virus was still there, not to mention the damage it had already done couldn’t be reversed, it had to be treated on day at a time. Norm said your lungs were getting better and your heart was too, but your kidneys, he thought they were called, still not cleared your blood properly, so you were still stuck in bed with a big tube coming out of your neck where it removed your blood and another tube where it returned it back to your body. Neteyam did not understand how any of this worked, but whatever science it took, he was grateful for. He did not leave your sight in this time, refusing to be parted with you for even more than a few minutes at a time. Jake helped Max and Norm bring in an extra large bed, like the ones made for Avatars and add it to the room next to your own bed, and he slept there, reaching over to hold you hand at all times.

There was so much between you that was still unsolved, still eating at his insides, but he did not want to overwhelm you or pain you in any way while in this state, so he was satisfied to just be near you and listen to your heavy breathing while you slept, and be comforted by the sound of the ECG machine letting him know you were still there, that your heart was still beating and it wasn’t stopping anytime soon.

“Ok, I think we need to start with how will we extract the proteins from these plants. I’ve not done protein extraction on plants, but I assume we can do some sort of chloroform or methanol precipitation? You and Grace used to work with plants a lot more than Max or I, so I think you would be the best at figuring it this out, no?” Neteyam looked at you with wide eyes, so perpetually impressed with your intelligence and drive, with how good you seemed to be at tackling any challenge. You were barely breathing properly, but here you were, trying to find a cure for this illness that almost took your life. The first full sentence you said after coming back to life was to Kiri, and he was a little jealous, but overall incredibly proud of you and your strength that felt like it could move mountains if you put all your might into it. Knowing you, he was sure this virus will be dead meat soon.

“Right, I can take care of the protein extraction, you can then run mass spectrometry on it and see what we’re working with. It’s going to be a lot of work, are you sure you don’t want us to take care of it? Until we figure out exactly which active ingredient in which one of these plants gets results, it’s nonstop. I don’t think you’re ready to be back to work yet.”

“I can’t sit back and do nothing. We’ll see how I feel, worst case scenario I do all the analysis from the comfort of this bed, deal?”

Norm was happy to hear that you weren’t being stubborn for once. Whatever happened to you when you died, you definitely seemed changed. As if death brought forth epiphanies that made you want to live, to heal. Neteyam was definitely not complaining.

After a while, everyone cleared, and you and Neteyam were left by yourselves. You turned around to watch him, giving him a little grin.

“You know you don’t have to stay here every minute of the day, right?”

“I disagree.”

You laughed heartily, your breath getting caught in your throat when the action made your lungs hurt, and the laughter turned to wincing, much to Neteyam’s dismay. Eventually, the wincing turned to sighing, and you didn’t look at him as you spoke again.

“Shouldn’t you be with your mate?”

It was his turn to wince, and he realised you probably couldn’t avoid talking about your issues forever.

“Do you really want to talk about this right now?”

“Neteyam… I won’t be the other woman. And I’m also not going to be the woman who steals a man from another, or who endorses whatever the hell this is. I need to make sure you are serious about this, about us. I might still not make the consciousness transfer. I need you to think about this, and if you do and you decide that it is me after all, then you need to talk to her. She deserves that, she deserves better than what she’s been getting. And I don’t want to be part of this until I know she is not being deceived or led on.”

“You are right. I will talk to her today, alright? But I’m not leaving you right now.”

“Do you want to watch a movie?”

You could feel yourself getting stronger by the day. It was a struggle still, your lungs feeling like they were going to explode any time you took a deep breath in and your heart oscillating between going a million miles a minute to barely functioning in the span of a day, but you were getting there. The combination treatment was definitely doing its job, and your body was no longer on the brink of collapse. Some colour returned to your cheeks in time, and since you have been eating about 15 meals a day because of how worried everyone was, you were gaining weight back, which was further increasing your day-to-day quality of life. You got a lot of sleep in the last few days, probably more combined than in the past few months, and you felt rested, for the first time in a really long time.

The first two days were the hardest, your body reeling from withdrawal on top of everything else you were experiencing. But you were feeling good today, and were trying to remain optimistic so as to not fall into old patterns. Having Neteyam next to you helped, your light in all the darkness, he refused to leave your sight, even in the face of more pressing matters.

The humans were closing in, you were told. The ships will be landing any day now, which only contributed to your slow recovery, as the anxiety was manifesting itself physically and hurting your progress. You felt stressed at the situation and more so at the thought that Neteyam wasn’t there, in the meetings and in practice, missing it all so he could stay with you. You hated it, worrying constantly and he wouldn’t hear it.

“It’s only a couple of days, Atan. Stop worrying.”

Today was the day they were going to remove the dialysis machine, which you were extremely excited about. You felt the need to walk, to stretch out your legs, to be back in the lab. You had a lot to do, a lot you had riding on this. You were trying not to think too hard on how you were doing everything in your power to further procrastinate healing, knowing that at some point you were going to have to deal with the trauma head on and open that damned drawer, that held all of your darkest memories, all of the things you spent your whole life pushing away. If you were going to be ready to heal, ready to move on, you had to do so with a clear conscience and a clean slate. You were adamant that you were going to find the cure for the virus and use it on yourself before you made a decision about the consciousness transfer. You had to be 100% sure of your choice, of your decision to leave this life behind forever, and you couldn’t do that if you viewed it as an escape from your current weak and disease-ridden body.

It took a while to be disconnected to the machine and get the tube out of your neck. From now on til you decide whether to go through with the transfer, you will have to do this for a few hours a day, which was a pain, but better than being bed ridden all day.

You put your feet on the ground for the first time in days, and you forced yourself to stand slowly, and quickly realised it was a bad idea, and your knees immediately buckled under you. Neteyam caught you with both his hands under your arms, and effortlessly raised you until your feet were a few inches above ground.

“Put me down! This is so embarrassing.” You groaned at your own impairment, and refused to give up, slowly starting to walk with shaky feeble steps. You felt both Norm and Neteyam hover over you, and although you were grateful for them, you wanted to scream at how upsetting it was making you having to be looked after like you were a baby taking her first steps. You guess, in some ways, you were.

You eventually got to your room, which looked untouched, if a little dusty. You were happy to see it again, happy to take it all in. It was a bit surreal, the whole experience. You didn’t realise how much it was all going to matter, all the little things, until you were dead on a hospital bed, begging Eywa for another chance. Every wall, every creak on the floor, every book, it all hit you suddenly, and you had to sit down on the ground, your back against the bed frame, so as to not collapse under the weight of this whole endeavour.

“Atan, are you alright?” Neteyam’s voice was laced with concern, and you tried to stop panting long enough to answer him.

“I’m fine, it’s just a lot.”

He slowly sat down next to you, struggling to squeeze in the tight space between the bed and the desk. His knee was brushing up against the side of the thigh, and you felt heat spread from the touch through your entire body. He gently raised one hand to the side of your face, removing strands of hair that were falling on it and tucking them behind your ear.

“Talk to me, Atan. I want to know, I want to help.”

You felt tears pool in your eyes, and you couldn’t look at him, couldn’t look anywhere but the ground as the words got lodged in your throat. This was going to be much harder than you thought.

“I’m so sorry. I’m so sorry, Neteyam. I’m sorry that I almost died, that I did die, and that you had to witness it. I’m sorry for not telling you, I am sorry for not fighting harder, I am sorry for so many things.” Hot tears were falling on your arm laying in your lap and you struggled to find the words to speak. “I am still so mad at you, so mad that you left, so mad that you lied, so mad that you never told me how you felt about me, so mad about so many things, but I shouldn’t have kept this from you. We’ve been inseparable since birth, and no matter what happened, you deserved to know, you all deserved to know. You deserved to say goodbye. I am sorry I took that from you.”

You were both crying now, and he picked you up and placed in his lap with little visible effort, it was probably easier for him than carrying Tuk. You wrapped your arms around his neck and slid your legs around him to wrap around his back and you stood there, crying in the crook of his neck while he held you tightly, trying not to hurt you.

“I am sorry for leaving. Sorry for not talking to you about how I felt, sorry about giving up on you. I should have stayed and helped you heal, I should have been there for you to rely on instead of being yet another thing you had to deal with in this life. I am sorry about lying about Tiongli, I am sorry for not telling them all no from the beginning. Even without the Avatar, there was only one woman in this whole world I would ever belong to, and I would have been happy spending my whole life in this lab, if it meant doing it with you. I was scared, and I am sorry.”

He removed your head slowly from the crook of his neck, and grasped the side of you face with one of his hands, thumb trailing over your lips, tracing them from your cupid’s bow to your lower lips, and you shuddered at the touch, deep desire pooling within you.

“I love you so much. I have loved you my whole life, from the moment I was born, and I will love you until the moment I die. I am so sorry it took me so long to grow, but I am ready now. Whatever it is, I will brave it with you, and for you. You will never have to do this alone again.”

You were sobbing now, unmoored at his confession that you have waited for your entire life. You were so overwhelmed by the love and affection you felt for this man, for the appreciation of knowing he was willing to hear the deepest, darkest parts of your soul and share in that pain willingly.

“I love you too. I missed you so much.”

After crying it out on the crook of his neck, letting your human hands caress the soft blue skin of his back and chest while he held you, fingers running up and down your spine in a comforting motion, you eventually got up from his lap slowly, and crawled on all your fours to your messy old desk. You took a key that was taped to the underside of your chair, and, with a deep sigh and tired eyes, unlocked the bottom drawer of the desk. You took out just one thing. A bracelet. You smiled at the sight, and tightened it around your arm multiple times so it fit on your human form. You looked at Neteyam coyly, giving him a soft smile, and you saw his hand reaching for his own bracelet, with a gummy smile and tears in his eyes.

Maybe you could do this after all.

You eventually started working in the lab again, being a lot more careful about striking a balance between work and everything else in your life. You were still struggling to sleep, especially now that Norm and Max flushed every pill you had down the toilet, but you were trying your best. The best was when Neteyam slept over, and you just fell asleep flush against his bare body, whose warmth melted away all your nightmares and replaced them with much better, much lewder ones. You weren’t getting anywhere with your research, as Kiri found a lot of plants in the forest, and to do mass spectrometry and proteomics on so many proteins, and analysing them all was an impossible task in the time you had left. So in an afternoon, as you were hanging out with her, Lo’ak, Spider and Neteyam in the hub, guitar strumming in your hands, you had a lightbulb moment.

“Kiri, if you had to save one plant, just one on this whole planet, which one would it be?” Kiri was special. Whenever the rest of the kids argued over her heritage, making crude jokes about Grace and Norm, trying to figure out who her dad is, you always had a different hypothesis, that you never said out loud. Kiri was Eywa’s child, as much as Grace’s. She was the key to everything. Eywa was flowing through her, guiding her, and you believed she had a lot more up her sleeve than any of you knew, than even she knew.

“Hmmm, that is a hard question.” She looked deep in thought for a long time, and eventually it’s like a huge epiphany was released from her mind. “Pamtseowll!!”

“The Cat Ear plant? Are you sure?”

“I think so! I don’t know why that one, but that’s the only thing that came to mind, and when I thought about it, I just knew it was the right one! Why do you ask?”

You raised from the ground, placed the guitar on a bench next to you, and ran (more like walked at slightly increased velocity than normal) to her, kissing her forehead in a loud smooch.

“I’ll let you know!! Wait here!!”

You talked to Norm, telling him your hunch and asking him to treat a vial of the infected blood with the liquid extracted from the plant in question, that you already had in stock thanks to Kiri. It would take a while, but you were buzzing in excitement, feeling extremely positively about your idea and your new odds. You made it back to the hub where they were all waiting for you, and sat back on the ground, mischievously ignoring their curious glares.

“Are you going to tell us what this was all about?” Spider said, rolling your eyes at your antics.

“Nope.”

They eventually dropped it, and you sat together, talking about everything and nothing, trying to ignore the looming doom of the ships slowly making their way to Pandora’s atmosphere, and the deadly consequences that would follow them.

You picked up the guitar again, deep in thought. Before you could help yourself, you found yourself speaking.

“I saw my mum.”

Everybody stopped and stared at you in shock. None of them said anything, and you swore you could hear a hairpin drop for sure in this big hall, or your heart pounding in your ears.

“When I died…” you started cautiously, not even knowing why you started talking in the first place, but knowing it was too late to back off now. “…I woke up in this new place I have never seen before. Eventually, I realised I was on Earth, in my mum’s childhood home. There were pictures of her on the wall, with her parents, she looked young, and happy… and alive.”

“Her house overlooked the ocean, and I went and sat on the beach and she came. Like it was nothing, she just came to me, and sat me down, and watched me cry on her shoulder until I thought I was running out of tears.”

This bit was easy enough to speak about, now came the real problem. You swallowed the big lump in your throat and continued. “She told me she did the same things I did. That she had been sad, so very sad, ever since my father died, the kind of sad that never goes away, the kind of sad I am.” You couldn’t look at them, preferring the look of your guitar that you were holding on to dear life as you spoke, and saw small tears crashing into the strings, splattering everywhere. “She said she didn’t try… to stay alive, to heal. Just like I didn’t. She said she had a choice, and she chose to die, because the hurt was too much for her to bear. She left me, orphaned me, put me through so much pain I’m still dealing with a decade later.”

You heard small gasps and sharp breath intakes and the new information, but couldn’t stop to acknowledge it, needing it out of your soul as soon as possible. “I was so hurt, so mad at her. That she left, that she didn’t fight harder to be in my life, that she left me with so many scars because of her selfishness. And then it dawned on me… that was me. I did that to you. To all of you. And I am sorry. I am so sorry.”

They all slowly sat up from where they sat and gathered around you in a circle, and hugged you.

“We forgive you. We’re just happy you’re still here. We love you, we want you to know that you’ll always be able to talk to us. You’ll always have us. We’re a family. Sullys stick together.”

You felt each of these words deep within you, the forgiveness something you craved like air, and you realised how much it must have meant to your mum. You were happy you could do that for her. You were happy something good came out of all the misery.

Later that day, all the Sully kids left, and you went to your room, carrying your guitar in your hands, and found yourself again, sitting on the ground, reaching for the bottom drawer key and unlocking it with shaky hands and muffled cries. It was time. You reached inside it for a photo album, with old school printed photos on it, and opened it. Your heart was hurting so badly you needed to place your hand on your hand to try to settle it, settle the pain and agony you felt. The first photo was a photo of you and your mum in the medical ward, holding you tightly against her chest, looking really tired with the caption “you had just been born a couple hours ago, how crazy is that?”. The second one - a photo of you, as a tiny baby, only a few days old by the scribbling on it. Countless photos of you, in various stages of growth, in various positions, all captioned by your mother, who loved you more than anything else in this world and whose love was loud and clear in every one of these images. The tears kept falling off the laminated pages and onto your legs, and your anguished cries took over the music playing in the background. An image of you and Neteyam, in a crib, his body much larger than yours even as tiny infants, but you were holding hands, sleeping peacefully next to each other. An image of your mum with Neytiri and Jake, smiling widely for the camera. A photo of your mum in her Avatar body with all the Sully kids, her holding Tuk in her arms when she was just born. An image of her in the lab, mid laugh with a pipette gun in her hand - no one would ever have been able to tell how much hurt she was hiding underneath, how close she was to death. Finally, an image of her and your dad, taken by your dad, of them looking into each other’s eyes lovingly. The caption, blurred and blotched from dried tear stains, read “the love of my life, the light in all the darkness”. You closed the album panting, and brought it close to your heart, just holding on to it while you cried.

You were spent by the end, but wanted to push through, needing to get this done all in one go. You removed another thing from the drawer, a camera, the gift Jake have you for your birthday. You eventually sat on your desk chair, and plugged it into your laptop. You clicked on the only file on it, and gasped at your mum’s face appearing on the screen, smiling while adjusting the camera to centre her face. She started speaking, and you clung to every word, making sure her voice and words will forever be imprinted on your mind.

“My child,

I know it’s been a while since you have since my face.” she says with a small laugh.

“I’m making this tape for your 18th birthday, as this should be around the time when your Avatar is ready for you to use. I gave Norm and the rest of them notes and doodles and research and I think they can do it, my love. I think they can build you the ramps new life, one amongst the Na’vi. I am so excited for you, you have no idea.

Happy birthday, bunny. You will forever be the best and brightest thing I will have ever achieved. I am more sorry than you will ever know that I don’t get to see you grow up, see the beautiful, intelligent, caring and loving person you already are blossom into so much more, but I want you to know, more than anything, that I have no regrets.

I have lived my life exactly the way I wanted to. I have explored new worlds, I have contributed something good to the world, I have lived and I have loved deeply and unconditionally. I just hope you do, too. I hope this world doesn’t break you, my love. I hope it doesn’t snuff out your light, because in that case, I will have failed you, and I am so sorry.

I know you are sad, and I know you have probably been sad for a while, and I know this life is emotionally abusive, and it takes and takes and takes until there’s nothing left. But it’s also worth living. Because you can learn to take from it, too. You can learn to heal, and love and live, and replace whatever it took in time with even better fitted pieces. But in order to do that, you have to live, my love.

Even at 10, you have a tendency to keep sadness in, and don’t let it out. That’s something you got from me, and I’m sorry for that. I love that you got my eyes, and my addiction to music and my love for science and literature, but baby, I don’t want you to be like me in every way. You also have a tendency to isolate, and that, you also got from me. I’m hoping that will change in time, I’m hoping Neteyam, and Spider, and Lo’ak and Kiri, and eventually Tuk will be able to break through and allow their light to shine on the dark walls of this lab. But if not, I’m hoping this Avatar will.

I want you to know that it’s alright to hurt, and it’s alright to be scared and wonder if the path you’ve taken is the correct one. We all wonder through this life scared and confused and a little alone. But it’s up to us to want to make a change and it’s worth it, baby. I know opening yourself up to hurt is scary and hard, but it will also allow to love, to love fully and deeply, to give yourself to others and have them give themselves to you. There’s beauty in this world and this life, but you need to be brave to experience it.

Your dad and I love you very much. Enjoy this Avatar, and enjoy all that it can bring to your life. You are the brightest light of my life, and now it’s time for you to become another’s.”

You replayed the video until you were so spent, your eyes were closing painfully from the tiredness and hurt and tears. After that, you went to your bed, still clinging to the photo album, and fell asleep, to a song you hoped one day you’ll be strong enough to play yourself, but for now had to settle for the original.

I should've asked you questions, I should've asked you how to be Asked you to write it down for me, should've kept every grocery store receipt Cause every scrap of you would be taken from me Watched as you signed your name Marjorie All your closets of backlogged dreams, and how you left them all to me...

Norm came bursting through the door of your bedroom, waking you up in a panic. “What the fuck, Norm??”

“We did it.” He came to your bedroom and took you by the shoulders, shaking you. He screamed and jumped in the air.

“WE FUCKING DID IT, ACE!”

You jumped from the bed onto your knees, so you were almost face to face.

“WHAT? ARE YOU FOR REAL?”

“YES, OH MY GOD, YES!”

You jumped on him and he caught you, and you just spun around in your tiny room, laughing and screaming at the beginning of your new life.

Thank you, mum. For looking after me even after you’ve gone. Thank you.

You had to toil a bit with details, but by midday, you were ready to get the substance, now purified and diluted in appropriate medium for being inserted in a human body. You watched as Norm prepared, and held your breath as he injected it into your now healed veins. You didn’t know whether it would work, or how many doses you would need, this was all so new, it was kind of crazy you were willing to inject it in your body with so little information, but you trusted your mum, and you trusted Kiri, and you trusted Eywa.

You felt good. So good, in fact, you knew exactly what you wanted to do.

Neteyam was in the village, in a deep strategy meeting about how they were going to approach the upcoming threat looming over everybody. His mind was only half listening, the other half too busy with musings of you. What were you doing? Have you eaten? Did you take your treatment? He was worried about you after yesterday, and felt guilty he didn’t stick around for the night in case you needed him. He is pulled out of his train of thought when he hears a loud banshee scream, and everyone’s attention turns to the entrance of the tent, to which everyone rushes. When Neteyam makes his way outside, his mouth drops at the sight of you, beautiful and wild, free on top of your golden white ikran.

You were radiant, smiling widely at him while you removed your queue and your visors and brought your hand to your forehead and greeted everyone at the meeting.

“Hey, kid.” The Sully patriarch came over and gave you a big hug, a conflicted look on his face. “Are you sure it’s ok for you to use your Avatar?”

“We did it, Jake. We found the cure.”

His mother and father both gasped loudly at the news and hugged you tightly, and you held on to them with all your might, pouring all of your love and apologies into it. Neteyam was too stunned to say anything, pure bliss escaping his body in the form of silent tears, that he wiped immediately from his face - he was in a meeting with all the warriors, there couldn’t be tears.

“I’m actually here to steal the future Olo’eyktan, if that’s ok with everyone. I will bring him back in one piece, I promise.”

“Fine, but don’t be too late. I don’t want to have to use the radio, am I making myself clear?”

“Yes, sir.”

Neteyam saw you click your tongue towards where you were standing, and felt a ping of arousal at your sight, at your confident demeanour. You used to be such a wild girl before life got too much for you to bear. He hoped he’d get to see that in you again, just like he was now. His hands made their way to his mouth and formed a circle around it, and he screamed loudly for his ikran, which appeared quickly and landed next to yours. He connected the queue and both of you took flight at the same time.

You flew like that for a while, and he was mystified at the experience. He has spent so much of his time riding with other people, flying as much a chore as a delight, but nothing that will ever be able to top this. You were screaming, fiercely and ferociously, doing barrel rolls in midair on your beautiful banshee, laughing loudly and urging her to almost bump into him, to which he gasped in mock annoyance and returned the favour. He followed you to the Hallelujah mountains, all the way to a cave on one of the floating rocks. It was beautiful, and he thought it would probably be breathtaking at night. You both landed, panting from the adventure, and made your ways to each other after petting your ikrans affectionately.

His hands immediately found your face, holding you tightly to look deep into your eyes. Your eyes softened looking at him, and he could see himself in them, looking intense and troubled.

“Did you really do it?”

You raised you own hand and brushed an unwieldy braid from his face, gently tugging it behind his ear. He saw your tail swish enthusiastically.

“We really did it.”

Neteyam’s mouth immediately found yours, desperate for your touch, the feeling of relief washing over him and it was so intense this is the only way it could be manifested. He needed you, craved your touch, craved the closeness, craved to make up for all the time you two lost. You moaned into the kiss and deepened it, opening your mouth, inviting his tongue to meet yours and dance in the way that made you both weak in the knees. You put your hand on his chest and pushed him and it caught him off guard, because he tipped backwards and hit the ground with a painful thud. You smirked and raised an eyebrow, and he moaned at the sight. You immediately got on top of him, straddling his lap and pulling him by his chest piece until your lips met his again. Your hands went into his hair and his did the same, and you let out a wail when he pulled on it, making your head tilt backwards, giving him access to your neck that he attacked mercilessly, leaving trails of his presence all over you. You felt him, hard against your core that was dripping in arousal, and you thoughtlessly starting grinding against him, feeling your mind flooded with sensations you never wanted to let go of.

“Neteyam, we can’t.” It hurt you saying these words, it was taking every ounce of self discipline you had, which at the moment was almost none.

“Yes, we can. In fact, I think it would be irresponsible of us not to.”

You laughed, smacking his chest playfully at his words. You brought your hand to his face and caressed it softly, and he closed his eyes in bliss.

“I want to do this properly. I will be one of you soon, for the rest of my life. I want to do this your way. I want to be mated to you the Na’vi way. Then we can do whatever you want, however you want. Deal?” You raised an eyebrow at him, smiling mischievously.

“Only if you stop raising your eyebrow at me, otherwise I make no promises.”

You laughed again, and laid next to him with your head on his chest, hoping this moment could last forever.

“Hey, I have to do something, and it won’t be pleasant. I was going to go alone, but I’ve made a promise to not go about everything alone. So if your offer still stands, would you like to come with me?”

You felt Neteyam’s hand stop caressing your hair like it had been for a while. He starts getting up, pushing your head off his chest slowly in the process. He looked serious, concerned, but he had a calming smile on, and when he took your hands in his and held them tightly, you knew you were gonna be ok.

“Let’s go.”

You got on your ikrans and flew back to the forest, feeling comforted by the knowledge he was there, next to you. You didn’t have to do this alone. You landed in the clearing next to the exo suit that belonged to you dad. Your heart was picking up pace from the sight, but it was time. It was time to work through it.

You slid off Neyn’s back and made your way to the suit, running your hands over it to remove some of the vines and eventually reaching for the dash cam. It was probably long dead, but you had to try, especially after what your mum said. You fiddled with the screen for a while, and eventually, it turned on, making you jump out of your skin.

In the view of the camera stood a man, tall and bald, wearing military clothing and reloading the guns that came with the suit. You’ve seen him before, in a photo in one of the jackets you found rummaging for clothes to wear. His name was Lyle Weinfleet.

“Captain, I think if we go through this clearing, we should be closing in on the tree of souls in about 3 clicks.”

“You’re right. But I don’t think we should do it.”

Lyle looked stunned, and he stopped whatever he was doing to turn around and look and your dad. “Barlowe, what are you talking about?”

“This is wrong, Lyle. You know it, I know it, everyone here fucking knows it. And if you don’t, you’re even a bigger psychopath than I thought you were. We have to go back.”

“I’m not fucking going anywhere, Captain. These freaks are going to kill all of us if they get their way. Someone has to stop them.”

Your dad picked up his gun and pointed it at him. “Are you disobeying direct orders, lieutenant?”

“You’re not my squad leader, Captain. And I have orders from above.”

A fight ensued which eventually resulted in the destruction of both the exo suits. Your dad got out of the suit, and the last time the camera picked up was Weinfleet running at him with a knife.

You lowered the screen and sat in silence for a long time. Neteyam’s hands were rubbing up and down your arms, and you felt his chest radiating heat on your back. You were happy to have him here, as the new influx of information was overwhelming enough to make you dizzy. Your dad didn’t attack on that fateful night 19 years ago. Everything you believed your entire life was false.

“My dad was not a horrible person.” Neteyam said nothing and just stood by your side, allowing you to process everything at your own pace.

“I hated him my whole life. I felt so much guilt at his actions, the murders I thought he committed plagued my mind for years and years. I was too scared to come to the village because I thought he was responsible for the decimation of your people.”

“You came from him, you share his blood. There was no way he could have been that bad. Not with you as his daughter.”

You scoffed lightly at his comment, but were so happy he was here. You took him by the hand and walked him to the body that was now only a pile of bones. You looked further in the clearing, and realised another set of bones was settled in the ground a few meters from your dad’s. Good, you thought.

“I’d like to bury him, if that’s alright with you?”

You gathered all the bones and walked to a tree whose roots were hanging partially above the ground, creating a little cove. Rays of light were hitting it, and you knew then it would be the perfect place. You placed his remains there, and gathered flowers and ornate twigs that had fallen on the ground, and decorated the little space as well as you could.

“Thank you, dad.” You were comforted at the thought that, although he died on this foreign planet alone, he was loved, and now, he will be mourned.

You made it back to the village together, walking and holding hands, trying to get your mind off all the epiphanies you have had to undergo in the past few days. Healing was hard work, you snickered to yourself. You made it to the tent you knew Jake and the rest of the warriors would be.

“There you are. Was starting to get worried.”

“Ah, there’s nothing to be worried about, pops.” You laughed at Jake’s shocked expression and the way his eyes softened immediately as a response to your new nickname.

He had to wipe that expression off his face. He was Olo’eyktan. He was Toruk Makto, this was not inspiring leadership.

“Right. The humans will be here any day now. I think we will have to move our base of operations in the Hallelujah mountains, somewhere they can’t find us, where the flux is too strong for their equipment. It’s going to be a big ask, and a big adjustment, but it is necessary. You know the first place they will look is at the hub. And it won’t take long for them to find the village either.”

What he was saying made sense, but it filled you with a sense of grief. This forest, that hub, those labs, this village, is all you’ve ever known. To have to leave it for an undetermined amount of time, maybe forever, was hurting your heart. You knew Neteyam felt the same, you knew they all felt the same. Jake was a marine, strong and adaptable, he was an incredible person to have around in a crisis. But he didn’t quite understand what it means to belong to only one place, to only know one reality that was going to be uprooted and taken from you. That takes time, time you didn’t have.

“I think we should do the ceremony tonight. You have completed the Iknimaya, you are to be one of us now. It’s time, if you want it.”

You peered up at him, in shock, but the beating of your heart was so loud because it was excited, it was happy. It’s time. You knew it in your heart it was right, this time. You belonged here. Your mum worked so hard for you to get to do this, she foresaw this 18 years ago. Your dad gave his life for yours, even without knowing it. You owed them this, owed them your life and this new one you were about to get.

“And tomorrow, we can do the transfer, if you are ready.”

By nightfall, you were ready. Your hair was freshly braided, in a pattern different to what the Na’vi wear, but you wanted it to be a mix of Na’vi and human braiding. You had all new clothes, and Kiri was in your tent, painting your body in white lines, as was the tradition. She looked radiant, happy and beautiful, like she always was.

“How are you feeling?

“Nervous.”

“There’s nothing to be nervous about. You belong here, you always have, even when you were human. This is just formality at this point.”

“Kiri… I want to thank you. You saved my life. Your plants, your mind saved my life. I wouldn’t do this if it wasn’t for you.”

“You saved your life. You asked me, you did the work, you found the cure. The Na’vi will be forever grateful to you, and Max and Norm, I hope you know that.”

“How about we say it was a team effort?” You laughed together and you kissed her forehead. You loved her so much, your sister for life.

Neteyam saw your body, adorning all new garbs, a golden frilly loincloth and green, violet and gold beaded top that hung from your neck to reveal a bare back that made Neteyam’s body beam with desire. Your left arm was decorated with a bracelet, the same bracelet he was wearing, the one that he gave to you when he was ten, that was now proudly shining on your person rather than hiding in a drawer, masking pain. You had bracelets around your ankles as well, that were softly clinking every time you took a step. Your hair was soft and braided in a pattern that was unlike the Na’vi ways, and it was fitting, Neteyam thought. You would never be only Na’vi - you were more, not quite human, not quite one of them, you were the most beautiful aspects of both and Neteyam felt his heart skip a beat at the sight, one which will never cease to take his breath away. You were his, forever. How did he get so lucky?

Almost as if you could hear him, you slowly turned around and he saw your eyes searching the crowd until they found him, and your face immediately broke out into a beaming smile, that slowly changed into a gaze filled with yearning and love….and lust. Neteyam gulped audibly at your expression, so intense it was making him crazed enough to almost consider taking you away right now, ceremony be damned, and coax screams out of you he would be sure anyone else would cower in shame at. But not you. You were his match, his twin flame, and he knew whatever he wanted to take from you, the screams, the pleasured pains, the moans and panted breaths, you would give willingly, and would take his in return, until you were both so spent you collapsed in aching bliss.

He waited though, despite his progressively harder to ignore twitch, knowing he wouldn’t, couldn’t rob you of this moment, knowing the waiting and anticipation would be worth it in the end.

The ceremony was ethereal, magical, and he couldn’t help reminisce about his own, years and years ago. The people all stood around you, waiting for the Olo’eyktan to say the words that will make it official. Neteyam saw his father approach you, wearing all the garbs worthy of a chief of the Omatikaya, and placed both his arms on your shoulders, smiling proudly. You were a Sully now, for all intents and purposes, and his father looked at you the same way he looked at all his children. Neteyam’s heart swelled at the sight, taking in the familial interaction.

“Ngenga 'ite Omatikayaä luyu set. Na'viyä luyu hapxì. (You are now a daughter of the Omaticaya. You are part of The People.)”

Neteyam made his way by your side, and he watched as the rest of his family did the same. He placed both of his hands on your body, one over your heart, and the other on your back. His family did the same, and he noticed tears in everyone’s eyes, including yours and his own. This was a moment none of them thought they’d get to see, that you never expected to experience, and Neteyam said a silent prayer to Eywa, thanking her for bringing the love of his life back to him, and into the clan.

After the celebration, you excused yourself from everyone, tired and dazed, as well as too impatient to wait for what you have wanted, needed for days, months, years. You found Neteyam casually talking to some friends, and you touched his back lightly, so lightly it was barely there. He shuddered minutely, and turned his body to face you. He took your breath away, always, but particularly today, in this light, under these circumstances, wearing his ornate clothes and that bracelet, the one that you would both wear for the rest of your lives. That one that meant forever.

“Come.” He said, and he looked at you like you were a meal that he’s been starved from, that he was going to devour. You bit your lip in excitement and followed him.

You went for a swim in a tiny lake not too far from the village. You swam together like that for hours, kissing and touching each other under the water, just feeling him. The anticipation was building strongly, and as the last of the paint on your body dissipated, you left, with hurried steps, quickly making your way towards the Tree of Souls.

Your entire body stalled at the sight. You have seen images of it, areal videos taken from drones and helicopters, you saw it all, and yet nothing could have prepared you for the beauty of this place, for its magnitude and significance. The air felt thick, charged with energy, and you could hear whispers all around you, soft, almost imperceptible, but they were there. Eywa was here, just like she was all around you, but here, you felt her stronger, almost palpably so. You made your way through the willow-like branches, seeing the purple hue illuminate your skin, and you just walked, taking it all in, touching it, experiencing it fully. You were so happy, it made you anxious. You’ve never felt happiness like this, and you were scared it wasn’t going to last, scared of knowing that the fall from such a high would break all of your bones, crush you forever. You felt Neteyam’s gaze on you, electrifying every inch of your body its gaze was touching. You could feel him, in your bones, you knew what he was feeling, what he was thinking. You shared a soul, you always have. “He is more myself than I am. Whatever our souls are made of, his and mine are the same.”

You looked at him and your breath hitched in your throat at the sight; he was the most gorgeous human you have ever seen. He was beautiful and angelic and looking at you like you were prey that he wanted to exert complete control over - and you didn’t mind. You wanted him so badly and you have waited for this for what feels like lifetimes. You knew looking at this man in front of you that you would allow him to ruin you, you would beg him to kill you softly, slowly drive out any ounce of sanity from your body orgasm by orgasm until there was nothing left of you but the echoes of moans you couldn’t help exhale. It was almost desperate, your need for him, and you heard your breath become laboured, panting with craving and ache. You saw him make his way towards you slowly, and stopped only when he was towering over you, and you had to bring your chin up to be able to look into his eyes. His hands immediately found your jaw, that he gripped tightly, making sure you weren’t going to look away from him, from that gaze that drove you mad and made you throb in pain, begging to be filled, craving sweet release.

“You’re mine. I can’t believe you’re mine. I have waited for so long, I have dreamt about this for years. The one thing I have wanted more than life itself and you are here. You’re mine. Forever.”

You were fully panting now, not even an ounce of shame on your features. You wanted him, you wanted him to know what he was doing to you. Wanted him to control you, to possess you, to take what was his, what has been his and will be his forever.

“Say it. I need to hear you say it, Atan.”

His grip on your jaw tightened, and you moaned, unable to stop yourself, not wanting to stop yourself.

“I am yours, Neteyam. I have always been yours. I will always be yours. It’s you and me, forever.”

It was his turn to moan, gaze turning animalistic now, in a way that you felt within you, deep within you, and you knew you were dripping wet now, knew he was going to love it, was going to revel in it, and you smirked at the thought.

You saw him stop and reach back for his queue. You gasped silently at the sight, but smiled at the thought he wanted this, that after this, you would be his for life. You reached back for yours, and you both held them close to each other, just out of reach, looking deeply in each other’s eyes.

“Do you trust me?”

“You know I do. I trust you, Neteyam, do whatever you want to me. I’m yours to take, I’m yours.”

“Forever?”

“Forever.” You joined the queues and nothing in the world could have ever prepared you for the overwhelming sensations and feelings, for the intense intimacy of feeling Neteyam’s deepest desires and emotions, for how deep his love and care for you actually ran, for how desperate he was to own you, to make you his. You felt his breathing and his heartbeat, and he felt yours and the desire you both felt exploded all around you.

He aggressively smashed his lips into yours, both of you moaning at the contact, at the need that enveloped both your beings like the rushing water of the river you loved swimming in together. Your fingers immediately made their way into his hair, his beautiful hair that seemed to sometimes have a life of its own, and thinking of how hot you found it made you only more enthusiastic, gripping at it tightly and pulling on it. He smirked in the kiss, parting your lips enough to look at you with a mischievous grin that you loved more than you could put into words. You loved this man - you loved his kindness and compassion and sacrificial streak and patience, but you loved this side of him more, the side only you ever got to experience, that no one else knew about. How passionate and adventurous he was, how masculine and possessive of what was his he was, how wild and untamed a streak he hid under the well-behaved and poised demeanour. How well he matched you. Your twin flame. He placed both of his hands on you thighs, just underneath your ass, and lifted you effortlessly, and you instantly wrapped both your legs around him, never breaking the kiss, that was now just a mess of panted moans, lips and intertwined tongues. You felt your back hit a tree bark and you gasped, breaking the kiss and watching as the action left a trail of spit in between you. His mouth moved to your neck, that he sucked and licked until it was hurting, and he pushed you into the tree, trying to remove whatever space there still was in between you two. As he did so, you felt his chest and pelvis push aggressively into you, and you whined when you felt his dick brush up against you core, hard as a rock and hitting a spot that was making you lose your ability to see around you. He started moving his hips into yours, drawing circles into your pelvis and you matched him, desperate for release, desperate to want to give him your first of many orgasms of the night. You weren’t going to be satisfied until he made you cry in pain, in pleasure, until the line between them was blurred.

Neteyam saw your breathing quickened and he smirked in a way that only made further pleasure pool in your depths. “Coming already, my love? You wanted me that badly, huh?”

“Dick.”

“You’ll get that later, too.”

You wanted to laugh, but couldn’t do anything but mewl in agonising ecstasy as he hit a spot that immediately made you see stars and you rode out your first orgasm, crying into the crook of his neck.

“Good girl.” He didn’t let you come down from you high, as he removed your body from the tree and placed you on the mossy, comfortable ground, and both his hands went on your knees, making sure you were keeping them apart, as soon as he could feel you wanting to close them to accommodate for your post-orgasm sensitivity.

“No, Atan. You wanted this, remember? You have to keep your pretty legs spread for me.”

The entire world was spinning around you, a mess of purple, pink and white, as he traced his hands slowly, too slowly, painfully slowly up your thighs, until he reached your hips, where you loincloth was tightened, and you felt him make quick work of the knot, inhaling deeply as he took it off from you and saw you for the first time.

“Fuck, Atan. I feel like I could come just from looking at you. Look at you, a fucking writhing mess and I haven’t done anything yet. Look at this,” he said as he place a hand in between your folds, removing the slick that was so abundant it was now dripping down your ass, “Look at how wet you are for me, and I haven’t even touched you yet. The shit I want to do to you, Atan, will make Eywa regret ever making this tree part of the mating ritual.”

You saw him slowly lowering his head towards you legs, kissing and biting roughly up your thigh until you were screaming in pain. “Shh, Atan.” He drew circles in your thighs as he continued his onslaught to comfort you to some extent, except it was only lighting your skin on fire, only strengthening your need for that second release.

“Fuck, Neteyam, please.”

“Begging so soon, my love? What do you need?”

“You. Your tongue, your fingers, anything. Fuck, please!”

“Well, since you said please.”

He smirked again, the asshole, and you felt him licking your pussy from your entrance to your clit, and at the moan he released as he did so, you convulsed so violently you needed a while to settle, still trying desperately to close your thighs to make the sensation more bearable.

“Stop that, or I’ll stop. Do you want me to stop, my love?” He looked at you through hooded eyes and giving you a wild look that made you clench around nothing, and your hands found his head and you pushed him back into you, annoyed that he stopped, already upset at the thought that he wasn’t going to be doing this every minute for the rest of your life. If you had your way, he would. That’s all you would do. He laughed, but obliged, and continued his attack on your lips, messily kissing and licking, pushing his tongue deep within you, like your pussy held the potion for eternal life and he wanted to live in you forever.

“Fuck, Atan, you taste so good. So good, how am I ever supposed to stop, huh?” You started grinding your hips around him, moaning loudly and you saw him dry humping the ground, trying to get some of his own release. The primal animalistic action was enough to push you over the edge again, coming all over his tongue, that was still lapping at your insides, without any intention to stop.

You tried to push him away, the feeling so overwhelming it was bordering painful, your clit so red and sensitive now it was glaring and obvious, but he didn’t relent.

“Fuck, you are intoxicating. Come on, baby, I’m not nearly done. We’re not nearly done.” He pushed two fingers in you, which was incredibly easy with the amount of slick and cum you were continuously releasing. Although so sensitive, you immediately clenched around him, taking him in, and felt the familiar ache slowly building in you again. He pushed his fingers deeper, until his knuckles made contact with your folds, and you felt his thumb slowly, gently rubbing your throbbing clit as his fingers were hitting your g spot mercilessly. The stimulation was too much, and you felt yourself seeking yet another orgasm, bucking your hips wildly against his fingers.

“That’s right, Atan. Let me see you come all over my fingers.” That’s all you needed to hear, and almost on command, you felt yourself coating his long digits with your cum. He removed his fingers from inside you and you cried out at the loss, but he didn’t care, too busy staring at his fingers, covered in the clear viscous liquid enveloping them. He brought both fingers to his mouth and he took them both in and sucked on them, and immediately kissed you, messily and forcefully, and you moaned at the vulgarity of it, and the taste of your own orgasms on his tongue, which was exploring your mouth like he had your folds earlier. You loved it, loved all of it, drunk on the feeling, on the connection you felt to this man that knew you, so well, so deeply, knew your body instinctually, like he touched you and had you all his life, and you squirmed when you realised that now you had a whole life to do this, every day, you would never have to live without this feeling again.

“Do you feel how good you taste, Atan? Fuck. I will never get enough of this.”

With one swift motion, he removed your top from around your neck and took its place, squeezing your throat until you couldn’t breathe anymore, slowly choking you. You threw your head back and arched your back, eyes fluttering shut, just taking in this feeling, of being completely owned, completely in someone else’s control. It was exhilarating.

“Look at you. Look at how you give in to me, it makes me fucking crazy.” He squeezed harder. “You drive me fucking crazy. I have dreamt about fucking you for so long. So many nights, having to stroke my cock imagining it was your mouth instead, you have no idea how badly I need you. How entirely and completely yours I am. You own me. My soul and my mind. It’s only fair I own your body.”

The hand that wasn’t around your throat went to his loincloth, that he removed skilfully, letting it drop to the ground around him. Your eyes fluttered open to take him in, his glorious body and that magnificent cock, that was even bigger that you ever dreamt, and you struggled to gulp in the position you were currently in. Your pussy clenched fully at the sight, and you were drooling now, saliva pooling around your mouth that you licked off like a kitty looking at her favourite meal. His fist wrapped around the girth of his cock and he started pumping himself, and you were close to being done again just at the sight, never in your life seeing anything more sensual, more erotic. You needed him inside of you now.

“Fuck, please, Neteyam. I need you in me.”

“I know, Atan. I need to be in you. I need to feel you milking me, like the good girl I know you are.”

With a couple more strokes, he lined himself at your entrance and slowly, deliberately, agonisingly pushing in, one inch at a time, stretching you deliciously, fully until he bottommed out, and you could feel him hitting your cervix, you could see his bulge in your lower abdomen. It drove you to the point of madness.

Your hand went to his ass, trying to push him even further in, despite not thinking it was actually possible for him to go anywhere else, but it was still not enough, never enough.

He removed your hand and gathered both of them and pushed them above your head, pining them to the ground with enough force that you couldn’t move them anymore.

“No, Atan. You don’t get to dictate this. We go at my speed.” And with those words, he started rutting into you brutally, giving you no time to adjust to him or to the pace. It was rough and wild and out of control, everything you have ever desired in your darkest, best fantasies. It was so dirty, so wrong it made your insides squirm in pleasure, in uninhibited bliss.

He maintained his pace for so long it was enough to steal two more orgasms from you. You were on the verge of tears, now, so overstimulated you didn’t know if you were feeling pleasure or pain anymore, but you didn’t care, you wanted more, always more of this, more of him.

“Fuck, Atan, I can feel you squeezing me again. You think you can give me one more?” You nodded weekly, trying to match his ruthless pace.

“I want to come in this pretty pussy. You want me to fill you up, Atan?” You could only moan at the words, unable to formulate any coherent thoughts or sentences, anymore. You were way past that. Your breath quickened rapidly and you knew you were really close, and he could feel you too.

“Yeah, that’s right baby, come all over my cock. I want to feel you come undone.” Those words were enough to rip another explosive orgasm out of you, and your clenching milked his cock dry, feeling thick spurts of cum fill you up, painting your pink walls white.

He collapsed on top of you with a thud, still buried deep in you, and you couldn’t help laugh exhilaratingly at his reaction, at what just happened, at the fact his was all real, not just an out-of-reach fantasy.

Eventually, he rolled from on top of you and stretched out an arm for you to cuddle up to him, which you did, feeling grateful for every moment, grateful for the intimacy.

You were both still panting, still recuperating and you lay in his arms in peaceful bliss, trying to catch your breath.

“Thank you.” You said playfully, drawing circles on his chest.

“What for?”

“For being an even better lay than I always imagined you to be in my dreams. Although I’ll find it hard ever being a productive member of this clan again going forward.”

He threw his head back and laughed, and the sound filled the forest and your heart, so relieved to be able to hear it again, so immensely grateful to get to experience these feelings in your life, that didn’t end prematurely no matter how many bad decisions you have made.

“You’re very welcome, Atan.”

You peered up at him bashfully, and you knew then you did at least one thing right in this life.

You fell asleep in his arms, and woke up in the link room, wide smile on your face. Max was waiting for you, and he returned your smile when he saw how happy you were.

“You look happy.”

“It’s just been a good day.”

“Yeah, I’ll bet.” He looked at you playfully. “Do me and Norm have to give you the talk?”

“Shut up, Max. In a completely unrelated subject, though, can I have some birth control from the medical ward, please?”

Your body was weak, and you needed a while to adjust to being back here after a whole day in your other body, soon to be your only body. You felt weightless, despite the pain, so happy and so in love. Your knees were weak from the memories, and you felt a bit empty at the lack of touch, craving him again, even though he’s taken so many orgasms from you, you knew you were going to be sore walking back to the village.

You went looking for Norm, who was already waiting for you in the medical ward, ready to give you a new dose of the injection and take some blood to see how everything is looking.

“It was a beautiful ceremony last night. Did you enjoy yourself?”

“Yeah, definitely did. It was incredible, I have never felt anything like it before.”

“Are you ready for tonight?”

“I think I am. I opened the drawer, little by little, Norm. I think it’s time. I am ready to take my place amongst the people, I am ready for my body to belong, I am ready to not have to worry that this virus will turn around and kill me at any point. My mum and dad both sacrificed so much for me to have a chance at a better life. You guys sacrificed so much. You’re like my guardian angels. I owe you so much, and I am so sorry about how I acted. I hope you can forgive me.”

“Will you still come by and hang out once in a while when you’re not busy riding your ikrans and being an overall badass?”

“Always, Norm. I am still me, I am still a scientist at heart, and I am still human. Maybe my body won’t be, but I will also have my mum and dad, you and Max, a whole history and heritage that I don’t want to rid myself of.”

“Good. Then all’s forgiven, Ace. I am sorry too, for not telling you about Neteyam. It just didn’t feel like my secret to share.”

“I understand.” You got up after he injected you with the treatment and hugged him tightly, grateful for this man who is your family just as much as your mum and the Sullys are.

After resting, eating and taking a nap, you did your dialysis session for the day, and although loopy, you hopped in your Avatar body, still a couple of things you wanted to take care of before the transfer.

To your surprise, you woke up in your tent, a small blanket covering you. You smiled, realising Neteyam must have carried you back in the morning. He even put your clothes back on. You blushed at the need that immediately overwhelmed your body, and tried to calm yourself as there were more pressing matters at hand.

You made your way to the Sully tent, and heard Neytiri and Mo’at talking. Good, two of the people you were looking for.

“Mai’te! How are you feeling?”

“I feel great, sa’nok (mother). Thank you for last night, it was beautiful.” Neytiri brought her hand to her mouth in slight shock at your new name for her. She was so happy, it all felt complete now, after all this time.

You took her free hand in yours as well as one of Mo’at’s, and looked at them seriously, intently.

“I want to ask for your permission for the transfer. I realise a lot of talk has happened, done when I was a human, when I was sick. But I am not sick anymore. I don’t want you to do it because you are worried I will die. I want to be sure, I want you to be sure, that you want me as a Na’vi forever. That you are comfortable with me becoming my Avatar for life. I will not do it, if you aren’t. This means so much for me, and I know I have been a coward, for so long, but I am ready now. I am standing here in front of you, and I am finally ready, to be one of the people, to be in this body, to be your daughter. I just hope I’m not too late.”

They both looked at each other, and smiled, and looked back at you lovingly.

“Mai’te, we have waited for you all of our lives. You are the missing piece in this world, in our world. You brought forth a cure for an illness that has plagued the Na’vi for years and years. There is nothing we would love more than for you to be Na’vi. And there will be no better future Tsa’hik.”

Your eyes widened at Mo’at’s words, at the consent not only to join the clan, this family, but to do so as Neteyam’s mate, as the future spiritual leader of the Omaticaya.

“I won’t let you down.” You hugged both of them, tears in your eyes.

You eventually made your way out of the tent and looked for the only other person you had to talk to before this was all done. Neytiri told you where he could be, so you looked there first. Sure enough, you found Lo’ak deep in thought up a tree, above everyone else, and you climbed easily and joined him. He was startled by you, but he composed himself quickly and gave you a look that you couldn’t quite place. You could tell he had conflicting feelings, happy on one hand to not have lost you in one way, sad that he did lose you in others.

“Hey.” He was curt, and didn’t look at you, and a twinge of hurt settled in your chest. You had to make this right.

“Hey, you. Scooch.” He moved slightly so you could fit with him on the branch, which was more than wide enough to accommodate both of you.

“Lo’ak… please talk to me? I can’t stand the thought that you hate me. You’re my best friend. Please?”

“Is it true?”

“Is what true?”

“I saw you leaving with Neteyam last night, after the ceremony.”

You cringed a little, the thought of having this particular conversation with him close to unbearable.

“Yes, it’s true.”

Pain flashed across his face and you saw a tear, just one small tear, fall down his face. You reached to wipe it off, but he beat you to it, so you awkwardly let your hand drop.

“I’m sorry, Lo’ak. I’m so so sorry. But I have loved your brother my whole life. My whole life, ever since I was born, he was there. And I love you too, I love you so so much, but we wouldn’t be right together. You are the best guy in the world, and I am not even close to being worthy of you. Neteyam gets me. He gets the darkest side of me, he knows my darkest, worst secrets, he has been with me through some of my most traumatic moments. It’s never been that it’s not you. It’s just that it can’t be anyone else. My whole life, it was always going to be just him.”

“You are my best friend, and I will always be here for you. You will never lose me. You will never get rid of me. I found a season of Friends on my mum’s directory that I’m pretty sure we’ve never seen. I can’t do that with anyone else.”

“I know you are scared, and I think that’s why you asked me. Not because you think I’m the woman you want to grow old with, but because you’re scared that no one else will see you the way I do. And you’re wrong, Lo’ak. Everyone will see in time that you are the best person in the world. You are incredible. And so many girls will be falling at your feet soon, you will be angry at yourself for ever pining over someone like me. I know you are scared, but I am not going anywhere. Can you please forgive me?”

He turned and looked at you for a long time, intensely searching your face, maybe for a sign that you were lying, or exaggerating, but he couldn’t find any, as he eventually dropped his gaze and smirked playfully.

“Do you really think girls will be falling at my feet?”

“Yes, I really really do. You are a catch, mister.”

He rolled his eyes dramatically and let out an exasperated sigh.

“Well then, I have to forgive you.” You both laughed and your back pressed against the bark of the tree and you stretched your legs to sit in his lap.

“Now, about that season of Friends…”

You returned to your human form in the afternoon, and slowly started packing your life away, ready to relinquish this room, once your mothers, with deep sadness in your heart. You will all have to relocate from tomorrow, so you will probably not be back here except to help everyone else evacuate and move all the equipment. You’ll never be back here, you realised with a deep sigh. So many memories, so many bad ones, but so many good ones too, this place was your safe place, your little piece of heaven, despite all the hell you went through in it. As you packed all of your books and trinkets you collected in time, you found yourself tracing your fingers over every surface, trying to commit it to memory. The coffee stain on your desk that was made when Neteyam spilled your mug with his tail without meaning to. The scratches on one of the drawers from all the times you pulled on it with your finger, since the latch was a little broken and hard to open. The wear and tear from your ass print on the chair that you spent inordinate amounts of hours in, be it analysing data or watching shows, or reading with your feet carelessly stretched on the desk. You moved on to your bed, that you sat on, and you sighed at the thought of never being able to sleep on a bed again. You will miss the comfort of the mattress and the sheets, and the space to really spread out however much you wanted, looking like a little starfish every time Norm woke you up in the mornings and his subsequent laughter at your ridiculous poses.

It took a while, but you were done about an hour before eclipse. Neteyam knocked on your door, with eyes slightly widened at the state of the now empty room.

“I’ve never seen this place like this. It looks so barren without your touch.”

You sighed and tugged at the insides of your mouth with your teeth. “Yeah, it does.”

“Any regrets?” You turned around to face him, giving him a small reassuring smile and taking his hand in yours, the size difference always a stark contrast compared to your Avatar body.

“Never.” You eyed the bed playfully and raised an eyebrow, feigning timidity. “We still have some time… what could we possibly do to fill it, huh?”

He looked at you, and you could tell he was turned on out of his mind, but also gave you a reserved look. “We can’t… not like this, I will hurt you.”

“I don’t think you realise you’re not making the point you think you are.”

He closed the door behind him with his tail and stalked towards you. “Fuck, you will be the death of me, Atan.”

You made your way to the village a little after eclipse, and everyone was following you. All the humans, Norm and the other Avatars and Neteyam, who was carrying you on his back like you were his little backpack. You were flushed and a little disheveled from earlier, purple marks all over your body, but it didn’t matter. Your hours in this body were limited. In your back pocket of your shorts you had a box of birth control pills that Norm gave to you as you and Neteyam made it out of the room. You laughed at his reaction and yours, both of you awkward and embarrassed to have to address it whatsoever. You placed your head on Neteyam’s shoulder, peering up at the sky, like you used to do when you were little. The village was busy and roaring, everyone packing their lives up for the move tomorrow. People looked at you as you passed by, unfamiliar with your human form clinging to the future Olo’eyktan’s back. Jake and the rest of the family met you at the big bonfire.

“Heads up, everyone. It’s time!” You smiled, and looked next to him on the ground, where your Avatar body lay motionless and you had to take a second to take this all thing in, the weight of what you were about to do hitting you full force. You started breathing in deep, shallow breaths, and you felt Neteyam’s hands running up and down your thigh in a calming motion.

“You’ve got this. We’re all here for you.” You reached over and pecked his neck, in gratitude.

Everybody in the village, all the Na’vi were making their ways through the forest towards Vitraya Ramunong - they would all partake in the ritual. Neteyam was nervous, out of his mind with worry and fear, although he didn’t want to impart that to you, so he carried you gently, trying to be strong, as he was sure your mind was also laced with concerns. What if you didn’t come back? Neteyam quickly pushed the thought out of his mind and cursed it for making him think about such things, making his heart constrict in his chest in pain at the mere thought. Of course you would make it. A part of him was also excited about the ritual. It would be the first time anyone under 19 would get to see it, to experience it, it was only the third time it would have ever been performed. It was a powerful ritual, hence why they needed everyone.

The tree was as breathtaking as it always has been, and Neteyam had to inhale deeper to accommodate for the deep charge of the atmosphere, for the thickness of the air. He heard the ancestor’s voices, buzzing quietly all around him. As soon as you reached the natural platform on which the tree resided, Neteyam put you down next to his family and tried to not think of all the images flashing across his mind’s eye of all the unholy things you two did last night right in this spot. His grandma was giving out orders about where to place your Avatar.

“Are you ready?” Said Mo’at, in her usual no-nonsense fashion.You gulped loudly, and Neteyam couldn’t help wince. Everybody was on the edge.

“Yes, I think I am.” He saw you turn around to face him and the rest of the family, as well as Spider, Norm and Max.

“This is it, I guess.” You grabbed his mother and youngest sister’s hands in yours, and squeezed, a small tear falling down your face.

“I love you, guys. Thank you for everything all of you have done for me. I am so excited to join this clan and your family, forever.”

Neteyam watched as you lay on the ground, naked except for some leaves covering you. He was going to miss you, he realises. This body is the only one he’s known you as for 18 years. He’s spent so may days and nights with you, with this small person who had so much in her, so much beauty and intelligence, so much hurt and pain, so much personality, so much fierceness and grit and so much care and empathy, he didn’t know where it could all fit in such a tiny body. You’ve grown up together, a human and a Na’vi, learning so much from each other, adapting together to the world around you and to each other’s mind and soul. He was going to miss this body, the body he fell in love with, the toothy grin and wild eyes so unlike anything he’s ever seen, your beauty marks that he used to trace with his finger and eyes.. and mouth, as of a few hours ago. You were everything, and have always been regardless of your difference in species, and colour, and height. He mourned a little, he realised, for the you you’re leaving behind, but was also incredibly thrilled to have you as one of the people, as his mate, mother of his children, Tsa’hik, for the rest of his life.

He hovered over you the whole ritual, unnerved at the way the tendrils were emerging from the ground and slowly enveloping you, until there was almost no bare skin left to see. His skin was covered in goosebumps at the experience, as the chorus of people asking Eywa to grant you life in a new body, undulating their bodies in unison, and his grandma’s words. It was unlike anything he’s every experienced.

Tìng mikyun ayoheru rutxe, ma Nawma Sa'nok. (Hear us please, Great Mother)

Pori tireati, munge mì nga (Take this spirit into you)

Srung si poeru, ma Eywa (Eywa, help her)

Ulte tìng ayoer nì'eyng hu ngeyä ya (And breathe her back to us)

Srung si poeru, ma Eywa (Eywa, help her)

Tivìran po ayoekip (Let her walk among us)

Na Na'viyä hapxì! (As one of The People!)

Srung si poeru, ma Eywa (Eywa, help her)

Eventually, the ritual came to an end when Mo’at screamed for everybody to stop. Neteyam reached over to your face, and removed your mask, kissing your cheeks and your hands, and saying a quiet “I love you” before he moved on, anxiously waiting to see if you would open your eyes. He let out a panted breath as you did, smiling tiredly at the family who was now jumping on you, making you choke slightly.

You laughed and his eyes found yours in the manic scene unfolding in front of him. You smiled lovingly and reached out a hand for him to grab.

“Hi.”

“Hi, Atan. It’s good to see you.”

You were sad as you were carrying your body, you tiny human body in your arms, and lowering it in the ground next to the Home Tree. You saw your family place a flower each in the little cove where you now lay, and you carefully redirected an Atokirina towards it, and watched as it landed on you, making its home in the little gap made by your foetal position.

You will miss this body, and all the memories you made in it. You will miss looking in the mirror and seeing your mother’s eyes and hair, you will miss being able to play her guitar. This body, however tiny and weak, was your home for 18 years, and it has stood by you no matter how many times you treated it badly or betrayed it. It kept you alive in a world that could kill you in an instant, and you were sorry to have to leave it behind.

“Thank you. Thank you, mum and dad, for giving me life and gifting me this body. Thank you for taking care of me, I know it wasn’t easy to keep up with this messy mind. Thank you.”

You turned around and fixed your gaze on your mate, the love of your life, the one that made it all bearable, and when met with his dazzling warm smile and his touch that set your soul ablaze, you couldn’t help be excited at the adventures that lay ahead of you, that you will always brave together.

-the end-

EPILOGUE

The man felt weak and dizzy, the entire room spinning and making him well nauseous. He didn’t know what was happening, what any of this was. He saw a bright light flashing across his eyes and he groaned in pain and annoyance at the way it was making his vision spotty. He grasped the hand of whoever it was that was doing this to him, and he was relieved when it stopped. It took him a long time to adjust to his surroundings, and was met with shock when realising he was in some sort of medical ward, surrounded by people in white coats. His gaze fixated on his hand, that was still wrapped around someone’s wrist, and couldn’t understand what he was seeing in front of him. Blue. A blue striped hand, that supposedly belonged to him.

“Captain, Captain Barlowe, can you hear me? You’re ok, you’re just disorientated. It’s gonna take a while to settle your mind, so take you time.”

He raised assertively from the bed he was laying on and made his way to the reflective windows all around him. He stood there, still, just taking it all in, finding it hard to understand, to process what was clearly displayed in front of the mirror. His body, in an Avatar.

“Captain Barlowe, due to your outstanding military record and previous acquiescence with this project, you have been chosen as one of the select few to carry out a special op on Pandora. You will be briefed shortly, but right now we have to make sure everything is in order.”

“I’ll be damned.”

For everyone who’s made it this far, I am so excited to announce the sequel series, The Archer, that will tackle all of the events of the Way of Water x coming to you soon 💕💕

Tag list (thank you thank you thank you x): @nuhteyam @eywas-heir @fanboyluvr @mashiromochi @puffb4ll @sassy-persona @simp4ff @mommyneytiri @inomoikawa @jackiehollanderr @jaysarchiv3 @meivap @dakotali @hlhl99 @eskamybeloved @erenjaegerwifee @winchestertitties @mommyneytiri @ultimatebluff @elizarikaallen @yeosxxx @ssc7514 @lolcaca @jackiehollanderr @bunnyrose01 @therealbloom @neteyams-queue @r1dd1kulus @whore4neteyam

@kikookii @iliyoo @velvetskies @rebeccao03 @im-in-a-pansexual-panik

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idle-daydreams · 1 year ago

Note

idk if you’re doing oc-reader requests. But uh…yeah

could you do a Yandere fyodor with a reader inspired by my oc, Nera?

the gist is, she’s the guardian of the book(the one everyone wants). However, she has a deep rooted loyalty to the agency. The only thing rivaling that loyalty, would be her love for Sigma, one of the few she views as family. She’s distrustful of people, but still nice to people she knows and likes. Her ability allows her to feel the emotions of others, but she’s very sensitive to it. She usually wears mostly back formal wear, liking pants better so she can defend herself if need be

[A.N.: Sorry for being so late, I've been busy with midterms. Hope you like it!]

Rat And Mouse

Tw: Yandere, kidnapping, manipulation, mentions of murder and torture.

Nobody knew about your connection to the Book.

It was something you’d never discussed - not with the agency, despite your loyalty to them; not even with Sigma, though he’d been born of its pages. You’d kept your head down, continuing to pretend that you were just another Agency member, just an ordinary person with a near-useless Ability. It was worth more than your life if anyone found out.

And yet somehow, Fyodor did.

You cowered before him as much as you could while tied to a chair. Though the room was cold, your skin was slick with sweat, hair hanging loose around your face. Your head was still a little fuzzy, but you were certain you’d been chloroformed in your apartment.

The man sitting before you smiled serenely, one leg crossed over the other.

“Be calm, dorogaya,” he said.

You grimaced. Your ability allowed you to feel other peoples’ emotions, yet you could sense nothing from him. Not anger, nor excitement, nothing. It was as though he wasn’t a real person, but an empty doll with blank eyes and an unsettling smile.

It was unnatural, inhuman, and utterly terrifying.

“What do you want?” you said, trying to keep your voice from quavering.

Fyodor spread out his pale fingers. “Come now, I know they do not hire idiots at your Agency. Do you believe I do not know about your connection to the Book?”

Your heart leapt to your throat. “I don’t know what you’re talking about,” you said, trying to inject as much confusion as you could into your voice.

Fyodor smiled. “How disappointing.”

He stood up. You’d never seen Fyodor Dostoevsky in person before. In photos he’d seemed unimpressive – a pale, thin man with lanky hair and a tired expression. But what those pictures had failed to capture was the malice of his demeanour, the aura of darkness that hung around him. It was almost palpable, making your flesh crawl as he came closer.

“Please,” you said. “I don’t know what you’re talking about. I’m just an innocent—”

“—secretary? Forgive me, but I do not believe you.”

You tensed as he placed a hand upon your face, expecting blinding pain before the oblivion of death overtook you. But nothing happened. His hand was ice-cold, but nothing else.

“I said to be calm,” he murmured. “I am not looking to kill you.” To your horror and confusion, he ran his hand through your hair in an almost gentle caress. “You are very beautiful. I should have noticed a piece of such monumental importance earlier.”

“I-I-” you began, “I’m not ly-”

“Ah, please do not. Sigma has already told me everything, so there is no need for you to lie any further.”

No. A leaden weight dropped in your stomach. Sigma wouldn’t have - he couldn’t have! ‘But he could,’ a treacherous voice in your head reminded you, ‘his Ability allows him to extract information from people with a touch, remember?’

You looked up to meet Fyodor’s indifferent gaze. “You’re lying.”

He shrugged. “Believe what you wish, dorogaya,” he said.

Then he turned and left the room.

You exhaled, mind racing a mile a minute. Sigma wouldn’t have betrayed you, even if he did have the ability to do so. He hated Fyodor, and had no reason to work with him ever since the Vampire Incident. But then, the only other person who’d known your secret was your ex – actually your first, and only – boyfriend, whom you hadn’t seen in several years, and whom Fyodor had no reason to contact unless he’d already suspected you of hiding something. And the only thing which would give him cause was...

‘No,’ you firmly told yourself. ‘It wasn’t Sigma. He would never betray me like that! I trust him with my life. Fyodor must have found out... some other way.’

You shook your head vehemently to convince yourself. It didn’t matter much at the moment anyway. What was more important was the fact that Fyodor would use you to find the Book. Though you were its Guardian, you possessed no supernatural link to it, only the knowledge of its location. Something that he could draw from you through torture.

You looked around. It was a small room. There was no window, no clock, no way to gauge the time. Fyodor had tied you up rather cruelly; the rope binding your wrists had been looped around your neck, so that you were forced to keep your arms bent at an unnatural angle to avoid strangling yourself. Your eyes welled with tears as the full extent of your plight hit you. Fyodor was a madman, a cruel and barbaric terrorist no matter how mildly he acted. Your ability was useless, your fighting skills non-existent - how were you going to get out of this alive?

‘Its fine,’ you told yourself. ‘The Agency will know that I’m missing. They’ll find me. I just have to hold it out till then.’

Your arms were burning, sweat beating your brow, when Fyodor returned.

“Ah, my bird has kept herself on her toes,” he said, as mildly as though he’d caught you in some trivial task. “I am glad my arrangement was not beyond your capabilities.”

“What do you want?” you said through gritted teeth. “I’m not telling you where the Book is no matter how matter what you do, so just forget about it.”

“No? That is quite the bold claim. Are you not afraid that I will hurt you?”

“It doesn’t matter.” You swallowed. “No matter how much you t-torture me or whatever. I’m not telling you anything.”

Fyodor nodded, looking only mildly disappointed. “I admire your resilience.” He reached for your hair again, running his fingers through it to straighten it out. “There is a lot about you that is to… admire.”

His words sent a chill down your spine. Fyodor movements were gentle, almost tender – more suited to a lover than to a lunatic. You tried to pull your head out of reach, only for him to pause with a raised brow.

“You do not like it?” he said. “Apologies.”

He stepped away. You followed his movements nervously as he began fiddling with the things on the sideboard. “Tell me,” he began, as there was the clink of glass, followed by the sound of pouring liquid. “Determined as you are to keep the location of the Book from me – what makes you think I do not already know?”

“What?”

You started. Fyodor turned back to you, a full wine-glass in his hand.

“Would you like something to drink?” he said, proffering the glass to you.

“No,” you said distractedly, ignoring the dryness in your throat. “What do you mean, you already know? You know where the Book is?”

Fyodor slowly took a sip of the wine as he strolled over to you. That faint, awful smile danced upon his bloodless lips as he leaned closer. “Well, we have used a page from it before,” he said softly. “Did you not wonder where and how we got it?”

Your heart skipped a beat. Your position as Guardian meant that you knew the Book’s location, but you had no way of knowing if someone had removed it from its place. If Fyodor had somehow found it…

“Ah, I have agitated you. I am sorry for that.” He held out the glass to you once again. The wine smelled enticing; it was a deep red colour, almost the rich red of blood in the dim light. Yet your stomach roiled at the sight of it. Fyodor had the Book… if Fyodor had the Book, then the world was already as good as done for.

You met his soulless eyes, and dropped your gaze to the floor.

“You’re lying,” you said miserably, trying to think of what to do next. It was your worst nightmare; to be battling wits against one of the smartest terrorists in the world.

“Why would I lie?” Fyodor casually circled behind you, tracing a finger across the rope around your neck. You braced yourself yet again, imagining his finger slicing your throat.

“If you already had the Book, you wouldn’t need me,” you said, biting back the scream that threatened to erupt from your lips. “You’d already be halfway already to your next awful scheme.”

“Oh?” He ran his fingers along the base of your neck, leaving a trail of goosebumps in their wake. “And what makes you think that you are not part of that scheme?”

‘It can’t be.’ Your heart drummed painfully against your ribs. The door before you seemed to dance, the shadows in the corners laughing at your misfortune. “You – I’m only as important as the Book. The only reason you could want me is because of the Book. There’s no other reason.”

“Before the vastness of God, everybody is insignificant. But for sinners such as us, there might be other reasons.”

Something simmered beneath your skin, a faint flicker of an emotion so dark, so twisted, it made you feel ill.

Desire.

Lust.

Obsession.

It disappeared in a flash, as though dragged back into the void.

“I will give you a chance to make your own decision,” Fyodor murmured from behind you. There was a soft swish, and the pressure on your neck and wrists slackened. You slowly pulled your hands to your lap, realizing what had happened.

“You’re letting me go?”

Fyodor stepped back into your field of vision. “I told you; I am giving you a chance to make your own decision.” He pointed at the door. “You wish to leave? Leave.”

You stared at him, alarm bells ringing in your head. “What are you planning? You’re planning something.”

He smiled that awful, soulless smile of his. “It would not do to reveal all of my secrets, yes? You must figure out some things for yourself.”

You looked from him to the door, then back. There was no point in arguing with him. Fyodor hadn’t become who he was by being honest or transparent. There wasn’t a chance in hell that this wasn’t part of some elaborate strategy – but this was also your only chance to escape.

You stood up slowly, joints stiff after the prolonged restraint. “I don’t – whatever you’re planning,” you began guardedly, as you began backing towards the door, “but you’d better drop it. I’m not so stupid as to lead you to the Book.”

“I wouldn’t expect you to be.”

“So—” You broke off as you bumped into the door. You were wasting time. Fyodor watched as you fumbled with the doorknob, making no attempts to stop you. His eyes – those empty, all-knowing eyes – chilled you to the bone. You looked away as you flung open the door, fleeing the room. There was a narrow corridor that led down a flight of stairs, but you barely paid your surroundings any heed in your desperation to leave.

‘I have to get to the Agency,’ you thought. ‘I have to tell them about the Book. And I need to go check up on the Book, see if it is still safe. It has to be – I’d have known otherwise!’

But an awful thought hit you just as you reached the front door. What if Fyodor did have the Book?

“What is it?”

Fyodor spoke from behind you. You jumped; you hadn’t realized he’d caught up to you.

“Nothing.” You spun towards him, hand still gripping the door-knob. “I – stay away from me!”

He raised his hands in a placating gesture. “I have done nothing,” he said. “That door is unlocked; you are free to leave. Why do you not do so?”

“Because you brought me here!” Wrung by a sudden jolt of anxiety, you threw open the door. A blast of cold wind hit you; it was night, a chill wind howling through the trees. A snow-covered landscape surrounded you, a foreign horizon visible from the distance.

“Where are we?” you said with dread.

“Siberia.” Fyodor cocked his head. “I did bring you here. But even I can make mistakes sometimes, can I not? Or do you consider me incapable of fault?”

“No! But—” Your head spun. You hadn’t drunk the wine; you hadn’t ingested anything back in the room. Were you still suffering the aftereffects of having been drugged? Or had Fyodor done something to you?

“But you said that I was part of your scheme,” you said, rubbing your temple. “Don’t tell me you realized your mistake in the last five minutes. So it stands to reason that letting me go is part of your plan.”

“Is it? What if you’ve already told me what I need to know?”

“Because I haven’t told you anything!”

“Haven’t you?”

“I—” You ground your teeth. You didn’t know. You didn’t know what Fyodor wanted. But if he was simply done with you then he would have killed you, which meant that letting you go was part of his plan. He was either hoping for you to lead him to the Book, or he was somehow using you to get to the Agency. There was no way to tell unless you wrenched it out of him.

You looked at him, and felt yet again, that faint prickle of desire radiating off of him. He claimed to be a human… and humans were capable of more than just fault. They could hunger, for more than just food or water. They could desire, and be swayed by it.

“You… want me,” you said slowly.

Fyodor smiled. “The awful thing is that beauty is mysterious as well as terrible. God and the devil fight; and the battlefield is the heart of man.”

“What is that supposed to mean?”

“Is it not obvious?”

It wasn’t. The only obvious thing was that he trying to trick you, to manipulate you somehow. Whether or not Fyodor was attracted to you – and the idea made you both want to laugh and scream – his main motive was the Book and the eventual destruction of the world.

‘I can’t stand against him,’ you thought. ‘If I try to foil him directly, he’d crush me. But if I stay close to him, I may be able to get a hint of his plans, enough to take down the Decay of Angels for good. I could just pretend that I’m f-falling for him or something. So long as I don’t reveal anything about the Book, it’ll be fine.’

It was a sickening thought. The very idea of staying close to Fyodor filled you with dread; pretending to be his lover was straight-up repulsive. And the longer you stayed with him, the more you were at his mercy.

But you were a member of the Agency. And you were a Guardian of the Book. And if that meant putting your life on the line, you were willing to do it.

You took a deep breath.

“You want me,” you said, fighting down the panic clawing in your chest. “That’s why you brought me here. And you’re letting me go because—?”

“I brought you here in a fit of passion, but it is obvious that you don’t want to be with me. And I do not want a woman to submit to me by force.”

“That’s considerate of you,” you said carefully. “But I hope it isn’t a lie. You really have no interest in the Book?”

His smile widened. “Even if I swear on my life that I am not lying, would you believe me?”

‘Fair point.’ “So if I walk away, you’d just let me go?”

Fyodor gestured towards the snow-covered woods in response.

You bit your lip. “And how far are we from the nearest airport? How am I to get home?”

“Are you really asking me that?” he said, raising a brow.

“I don’t fancy travelling through the snow at night by myself. Your guidance is as good as flailing around blind.”

He inclined his head. “True. Well, we are far from the nearest city, at least a couple of days on foot. In any case you would find it difficult to go home from there — you have no documents on you, after all.” He slyly cocked his head to the side. “My private transport will be here in a couple of weeks; you are welcome to stay with me until then.”

The revulsion deepened. But you forced yourself to nod politely. “Then… I guess I have no choice but to stay.”

“A difficult decision.”

“But—”

The words stuck in your throat. How could you talk about living with a murderous psychopath like him? How could you simply state your demands as though hashing out a deal with a roommate?

Fyodor stepped up to you, leaning to whisper in your ear. “Do not worry, dorogaya,’ he said softly, his breath tickling your ear. “I will not force you in any manner. Anything you do will be entirely by your own will.”

You shivered. His presence pressed upon you like a shroud. ‘This is just a game,’ you reminded yourself. ‘A game of cat and mouse. Just play along for a while and you can bury this guy.’

“Thank you,” you said out loud. “Could you please step aside?”

Fyodor pulled away. You threw a glance over your shoulder, at the stars twinkling in the night sky. Then, squaring your shoulders, you stepped into the Rat’s Lair.

#yandere bsd #yandere fyodor #bsd imagines #bsd x reader #yandere bungou stray dogs #yandere fyodor dostoevsky #manipulation

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katiajewelbox · 1 year ago

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A close up look at DNA extraction from Medicago leaves ☘️. The small plastic tube I’m holding shows 3 distinct layers. The tube contains pulverised frozen leaf tissue mixed with CTAB buffer and chloroform. This chemical mixture causes 3 distinct layers to form in the tube based on their density. The dissolved DNA is in the upper layer (the least dense) where I can collect it and transfer it to a new tube for further purification with isopropanol and ethanol.

#katia plant scientist #botany #plant biology #plant science #plants #biology #molecular biology #science #mad science #laboratory #labcore #sciencecore #women in science #eppendorf tube #sample #dna #medicago #chlorophyll

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instruth · 5 months ago

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An Extract Of Dementia

An extract of rich dementia

Let the bygones be bygones

Read the Book of Alzheimia’s

Step in, breathe in the Ozone

Intoxicated to the blur amnesia

Jump off the Cliff of Oblivion

Enroll in the class of extinction

Start off with forgetfulness

Revive the joy of happiness

Take a leap to the darkness

Don’t you want to remember?

Fire from snow, red from amber

Drunkenness from being sober

Falls or the White Cliffs of Dover

President from ordinary members

Sniff a doze of chloroform

Sleep on, in the dental chair

Monitored by a girl in uniform

Rub your head, count your hair

Tooth bent as worm, on golf fair

©Johnny J P Lee

13 June 2024

A Gogyoshiren Poem (20)

Photos: J. P. Album Collection

#poetryportal #writerscreeds #smittenbypoetry #spilledwords #writingthestorm #poeticstories #inkstainsandheartbeats #writtenconsiderstions

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mxwhore · 10 months ago

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the good thing about failing so many rna extractions is that im slowly building up a resistance to chloroform

#this bitch is becoming unkidnappable

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whocookedthelastsupper · 10 months ago

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“Thomas Jefferson in the nineteenth century wrote to his daughter that childbirth was "no more than a jog of the elbow," when his wife had died in childbed, as this daughter was to do two months later. Much more honest was the anxiety evinced by Madame de Sévigné, when her beloved only daughter suffered three pregnancies in the first two years of her marriage, including a severe miscarriage. In a furious letter she warned her son-in-law that "the beauty, the health, the piety and the life of the woman you love can all be destroyed by frequent occurrences of the pain you make her suffer" threatening him, "I shall take your wife from you! Do you imagine I gave her to you so that you might kill her?" Françoise survived this pregnancy, but her mother's fears were not at an end. Immediately after the delivery she dashed off a hasty warning not to rely on breast-feeding to prevent conception:

"if after your periods start again, you as much afarink of making love with M. de Grignan, you may consider yourself already pregnant, and if one of your midwives tells you differently, then your husband has bribed her!"

The position of the husband in this all-too-common situation, caught between the options of lethally selfish sensualist or reluctant celibate, was not a happy one. He would, however, survive his sex life: very many women did not. And as the modern age with its much-vaunted progress and prosperity broke about the ears of women in the West, they had the disconcerting experience of discovering that childbirth became worse, not better; for in one of the decisive power struggles that were to touch all women's lives, men finally won their long fight to take over the management of women in labor. Male attacks on women healers were nothing new —one facet of the witch hunts had been the determination of university-trained male physicians to eliminate the female opposition. But with the advent of drugs, obstetrical forceps, anesthesia and formal medical training, male practitioners were finally able to usurp women's age-old role as birth attendant and present themselves as the chief accoucheur.

Armed with the authority of the expert, the new men had no difficulty putting down the old women, even when they were horribly in the wrong. On his own admission "the great William Smellie," the mold-breaking "Master of British midwifery" when learning his trade once slashed a baby's umbilical cord so clumsily that the child almost bled to death. Smellie informed the suspicious midwife, whom his arrival had displaced, that this was a revolutionary new technique designed to prevent convulsions in the newly born. Privately, though, as he later disclosed, he was never so terrified in all his life.

With the advent of chloroform and disinfectant in the West, medical science began at last to make headway against its own darkest prejudices that the suffering and death of women in childbirth were no more than a "necessary evil" to be seen "even as a blessing of the Gospel," as one leading British gynecologist wrote in 1848. Elsewhere, though, it seemed that nothing could dislodge the fatalistic attitude toward the loss of female life, nor change the habits and customs that promoted those deaths. From India, a British woman surgeon, Dr. Vaughan, sent this despairing report in the last days of the Raj:

On the floor is the woman. With her are one or two dirty old women, their hands begrimed with dirt, their heads alive with vermin... the patient has been in labor for three days, and they cannot get the child out. On inspection, we find the vulva swollen and torn. They tell us, yes, it is a bad case, and they have had to use both feet and hands in their efforts to deliver her... Chloroform is given, and the child extracted with forceps. We are sure to find holly-hock roots which have been pushed up inside the mother, sometimes string and a dirty rag containing quince seeds in the uterus itself... Do not think it is only the poor who suffer like this. I can show you the homes of many Indian men with university degrees whose wives are confined on filthy rags and attended by these bazaar dhais.

With great clarity, Vaughan saw that the root cause of this suffering, infection and death lay not with the dhais who ministered to the women, but in the attitudes of the husbands.”

-Rosalind Miles; Who Cooked The Last Supper? The Women’s History of the World

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whumpees · 3 months ago

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Kyle xy whump list

Synopsis: it's a sci-fi drama about a mysterious teenage boy named Kyle, who is found wandering alone with no memory and no belly button. Taken in by the Trager family, Kyle struggles to adapt to the normal world while exhibiting extraordinary abilities, such as incredible intelligence and superhuman reflexes. As Kyle tries to uncover the truth about who he is, he encounters challenges that reveal he is far from ordinary. The show explores themes of identity, family, and the unknown.

Episodes: 43 eps (3 seasons 2006-2009)

Warning: the show was supposed to continue after season 3 but got cancelled and in the worst cliffhanger possible 🙂 but nice show overall i enjoyed it. Watch the interview where the producers talk about the ending to know where the story was supposed to go.

_______________________________

S01

Ep1: emotional over someone playing piano and cries. runs through glass, minor cuts on his face, wakes up nicole and tells her in a little shaky voice "I am leaking" aka bleeding 😂, Crying. Teary eyes

Ep2: emotional, teary eyed "looking for his family:("

Ep6: nightmare. emotional and crying

Ep7: nightmare

Ep8: thrown to the ground, put in a choke hold

Ep9: sudden headache while using his super senses. Wobbly after using his senses. Again feeling faint while using his senses, collapses and has an aggressive seizure, in the hospital. Emotional, crying

Ep10: chloroformed with a cloth and kidnapped, half awake in the car, unconscious on the ground, takes a bit to finally come to his senses. Teary eyed

_______________________________

S02

Ep1: knocked out

Ep2: headache while arguing with someone. Snd again later

Ep3: teary eyed

Ep4: falls and hurts his shoulder during extreme training, later putting cold/hot water bag on his shoulder. glassy eyes

Ep6: head pain

Ep8: head pain. And again

Ep9: inside an underground lab and breathing toxic gas, decides to go back into the lab knowing that he couldn't survive much longer in the gas, feeling weak and breathing heavily, (flashback: foss doing cpr on him) struggling to walk and leaning against the wall, makes it out but collapses on the entrance unconscious, amanda giving him cpr, wakes up

Ep11: kind of an explosion throws him away on the ground, just laying on a stretcher but is fine

Ep12: sways, dizzy, blurry vision. Again feels dizzy suddenly and falls on his knees, blurry vision, puts his hands on his head in pain. hearing everybody arguing by his super senses and gets overwhelmed and dizzy, head pain, blurry vision

Ep14: winces in pain. Strapped to a chair to extract data from his brain and told if he resisted it'd fry his brain, given medication through iv, information being extracted, resisting and shaking, scientists say "he's on the verge of cardiac arrest" but they keep going anyway. Is fine

Ep19: cut on his finger and it bleeds a little. Heals someone and then has nose bleed

Ep20: sleep walking. Nicole told him he's sleep walking because of stress and that he should make his life 'simpler' BUT literally EVERYBODY keeps asking him for help all at once he became so stressed and started being clumsy and not pay attention which is completely out of character for him, jessy asks him if he's been sleeping and that he looks worn down and that something is wrong with him, loses control over his powers, everyone got so concerned for him cuz he kept making horrendous mistakes and told him to get some rest. He'd wake up and find that he drew dozens of stuff and he didn't even realise he fell asleep. Turns out using his powers to heal that person is what took a toll on his body. Overwhelmed by sensory stuff and things are too loud for him he feels pain, holds his head in pain and uses his powers involuntarily and it causes a mini fire

Ep22: uses his powers to resuscitate jessy and says if he fails it could kill both of them, succeeds then collapses

_______________________________

S03

Ep1: electrocuted and passes out, kidnapped, half awake in the ambulance and muttering "amanda" then injected with a sedative i guess, wakes up strapped to a bed with iv on and hooked up to some machines, tries to free himself but can't rip the straps. Takes a fake bullet for someone and falls unconscious on the ground (he didn't know it was fake), woke up on the floor and some men sit him up on a chair and he's still trying to process what's going on and his vision is starting to clear

Ep2: nightmare and wakes up startled. Amanda getting kidnapped messed him up and made him so anxious about her he kepy having nightmares, she left his sight for 1 min and he freaked out and started screaming for her (her reaction to it was hilarious she got freaked out that he got freaked out and was like WHAT???!!! 😂😂😂😂). Suuuper anxious all day cuz the psychic told him something bad will happen to his soul mate

Ep3: crying

Ep4: depressed over amanda and everybody is trying to help him move on 😂. Super drunk (to forget amanda), clumsy and close to getting himself into trouble at the bar cuz of his innocence and lack of social knowledge (it was so cute "i love you" 😂😂😂😂) and blurting out all his secrets in front of everyone, swaying, helped to walk, says he feels really sick and throws up. Receives bad news and tears up

Ep5: tears up. On the verge of crying

Ep6: tries his invention and falls off of it

Ep7: teary eyed (cute)

Ep9: jessy had a nightmare and started choking him thinking he was someone else. Using his powers to resuscitate jessy and is in pain (faking)

Ep10: electrocuted, weak and struggling to walk, electrocuted again and falls on his knees. They threatened to hurt his family and WE GOT TO SEE KYLE LOSE CONTROL OVER HIS ANGER FOR THE FIRST TIME AND ACTUALLY HURT SOMEONE AND HE COULD'VE EVEN KILLED THAT PERSON IF IT WASN'T FOR THE PLOT TWIST!!!!!! "nicole had been right when she said she didn't recognise the person i had become" OMG I WANTED TO SEE MORE OF THAT the worst cliffhanger in history I'm devastated the show was cancelled after we finally got to see kyle lose his innocent side i wanted to see him capable of hurting people who don't deserve good. I want to see his ANGER and how living in this world will eventually turn him morally neutral and not that idealistic "good" they make all heros be

#kyle xy whump list #matt dallas whump #english whump lists

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drluciengaudet · 2 years ago

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To say Lucien took the news of Aurelius' death poorly was putting it mildly. After an extended internal disassociation and a short clipped exchange between his slave who wouldn't, per usual, do as he requested he reassessed the situation and what and who he'd need. Hence how he ended up where and with who he was. The climate of things as they were gave him no reason to cover his tracks. Who would blame him for taking the head of magical security with him to Egypt. Furthermore he had every right. Cairo was his. Granted, he would now have a purpose other than being an insufferable reminder of failure.

Lucien had gone to Egypt in hopes to find it was all a farce, some plan he couldn't understand laid out by his former sire. Faced with the ashes of Aurelius Hathor the lines between genius and madness started to cross as much as all the unsettled and unresolved moments shared between him and the strigoi. Lucien took the strigoi's ashes and sun ring back to his lab in France along with his son, their son.

He'd cornered Cairo there and demanded the impossible. Lucien was no expert at Necromancy, but he knew its limitations and understood a vampire didn't have a spirit, that Cairo couldn't bring Aurelius back as with necromancy and couldn't allow the strigoi's spirit to posses him eternally as a medium because there was no spirit left. With magic out, Lucien turned to science. The probability was so low, but Lucien would give anything for the chance to have Aurelius back. He had to.

While his son tried to compel him to return to the castle Lucien ended up chloroforming him and strapping him into one of his hybridization machines. There was a humane way to do these kinds of things, but Lucien had never been one to be humane to his test subjects and when it came down to it, that's all Cairo was and a failed one at that. Cairo was genetically a mix of himself, Raphael, and Aurelius so technically all Lucien needed to pull out was Aurelius genetic material and he could leave the rest and Cairo alive. This could also be done by the da vinci like operation machine in setting all the designated needles in to the bones for extracting bone marrow and spinal fluid, but Lucien set to doing each with no anesthesia or numbing of any kind.

Organs weren't really needed, but was going to take them out for good measure and had cut down Cairo's chest and was bout to split open his breast plate. The scalpel was still in his hand when he felt the unmistakable presence of his current sire. The only sound the pumps of his machines extracting the life from Cairo who had finally stopped screaming making Lucien realize the roaring noise in his head was momentarily quiet. The three piece suit he wore was coated in Cairo's blood, Lucien not even bothering to put on any surgical gear. He stepped in front of the gurney as if he could keep Raph from his victim.

"You have no right to be here, De Luca." Lucien growled defensively combative pointing the scalpel at him. "This is my lab, this is my space."

@nottheangel-raphael

#nottheangel-raphael #blame it on love

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icryyoumercy · 1 year ago

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@nimblermortal

first, because these things are important nowadays, i am not a medical professional, this is not professional medical advice, the WHO has helpful information about malaria, when living or travelling in a region where malaria happens, mosquito repellent and appropriate protective gear are mandatory and non-negotiable, and so on

quinine is made from tree bark, which makes it one of the two medically useful things made from tree bark i know of, and also makes me wonder how people learn these things. were they just. walking around biting random trees. is the desire to chew on tree bark just part of what makes humans human.

it has initially been used as a muscle relaxant by the quechua people, to treat uh. random shivering? which is apparently a thing people sometimes do? which i could look into, but then i'd probably get distracted

the spaniards brought it back to europe, as they did with so many things, and because things like germ theory and microbiology and chemistry were still centuries from being discovered, what people knew of malaria was that it causes fever and thus shivering with very noticeable periodicity. and they had just been told about a thing that can stop shivering, so might as well give it a shot. if the symptoms are all you are aware of, the symptoms are all you can treat.

and for some utterly baffling reason, it turned out it didn't just help against shivering, it actually cured malaria? which. wasn't what anyone was aiming for, but gift horses and all that.

rome, being located very conveniently in a swamp, and having a rather inconvenient amount of popes and other people important to the catholic church, was perfectly located to pioneer such treatment and make a great many of the rich and powerful (and thus by advertisment of word of mouth and rumor everyone else) want some more of this marvellous drug, which made quinine (that is, the bark it's extracted from) one of peru's most important stolen goods

then, of course, a lot of fucked up colonialism happened (including in africa, because it's hard to do colonialism while dying of malaria), because europeans were unwilling to engage in things like fair and equal trade with non-europeans, we get fun medical price gouging and attempts at monopolies and general unpleasantness, and someone finally managed to isolate the exact chemical compound instead of just grinding up the bark and mixing it with something that tastes better than tree bark

and around the 1940s, malaria treatments with fewer unpleasant side effects were discovered (which i know nothing about and won't look up because adhd), and by 2006 the WHO has declared that quinine shouldn't be used as the first choise in treating malaria for a variety of reasons, including resistant strains and aforementioned side effects

also, if you're really curious about the taste, tonic water is traditionally made with quinine, and has been used as a prophylactic against malaria. once it wasn't used for that purpose any longer, though, people have decided to add less quinine and more sugar and citrus because they didn't enjoy just how incredibly bitter that stuff was. also, the FDA says you can't have more than 83 ppm of quinine per liter of tonic water, so if you wanted to treat malaria with it, you'd need to drink some ten liters per day, and if you want to use it for prevention, you'd need around 20 liters per day, at which point malaria seems like the better option

what tonic water can help with, on the other hand, is muscle cramps! not sure how much of that is the quinine and how much is the placebo effect, but at that point, we're back to readily available and comparatively harmless

either way, in the 1860's, it was one of the few actually working medical things (along with chloroform and diethyl ether for general anaesthesia, and opium for pain relief), so they will throw it at anything that has even the slightest ressemblance to periodic fever (to be fair, a number of other things they did also had the required medical effects, they just ran afoul of paracelsus's basic adage of toxicity

Alle Dinge sind Gift, und nichts ist ohne Gift; allein die Dosis macht, dass ein Ding kein Gift ist. All things are poison, and nothing is without poison; the dosage alone makes it so a thing is not a poison.

—Paracelsus, 1538

by reaching the poisonous dosage at the same or a much earlier point than the therapeutic dosage)

#three cheers for thorough investigation #a study in sex drugs and french #there are many things i could and probably should say about colonialism here #but this is what i do for fun which colonialism very much isn't #it was bad it was wrong it shouldn't have been done #and someone much more patient and mentally healthy #needs to work out the reparations we owe #also of course i cannot resist bringing up paracelsus #the man was baptised Philippus Aureolus Theophrastus Bombastus von Hohenheim #any excuse to bring this up is a good excuse

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solo1y · 2 years ago

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Extracts from the Diary of Alan Watts

16th May 1965: Myself, Allen Ginsberg and Tim Leary have been trekking through the Amazon rainforest in search of the sahatasca, a perhaps legendary form of ayahuasca which enables a man to levitate and see through dimensions. Leary and Ginsberg have been tripping absolute balls since we got here, which they believe will help in our search, as if the sahatasca had a mind of its own and could sense its people. We stopped for the night under a large tree. My two friends refused to lie on the floor because they thought it was full of moving snakes. On this occasion, they were correct, but none of these snakes are dangerous.

17th May 1965:

One of the guides died last night after being bitten by one of the snakes. The other guide appears to be fluent in every language except English. He can expertly translate from one tribal language to another which is not much use to us. Leary is still tripping balls and insists he understands what the guide is saying but that I am speaking Chinese.

18th May 1965:

Ginsberg disappeared in the night. He left a note in Portuguese which I was able to translate using my high school Spanish. He claims that he got a psychic message from Bob Dylan and had to go on tour with him immediately. I have no idea how he will find his way out of the dense jungle or if he does, how to make his way from rural Brazil to Los Angeles without his passport, which he jammed into a coconut before leaving.

19th May 1965:

Ginsberg has returned. He keeps calling Leary “Bob” and insists that he entertain us all with a few songs. Leary was also tripping balls and would only sing as Joan Baez. This caused mass confusion which caused Ginsberg to pass out and our guide to apply a cloth gag to Leary’s face until he calmed down.

20th May 1965:

Our guide went missing for a few hours leaving me with an unconscious Ginsberg and a chemically sedated Leary. Our guide had to use enough chloroform to, in his words, “knock out a whale” to keep Leary down and even now he keeps twitching. I asked our one remaining guide how exactly he would have set about choloroforming a whale, to which he said it would be a piece of cake after wrestling Leary in the rainforest mud for twenty minutes. I don’t remember it being that long but I was occupied by a mushroom I found on a tree which turned out to be some sort of lizard.

21st May 1995:

I have travelled thirty years into the future. All drug have been legalised. As a result, there are no wars or crime of any kind. I will admit that there was something of a dip in productivity but there are only one billion people on the planet now as we are far more sensible about breeding. Leary has been completely forgotten but there is a huge statue of Allen Ginsberg in what used to be Times Square. He would have loved it.

22nd May 19,995:

Quatzo atcl derrr beep beep in smersh bod. Ment furgy smool alla party. Ohn smeft Tim Leary gurge Ginsberg haft larnack.

26th May 1965:

I have woken up in Dudley Road Hospital in Birmingham. It’s taken some time with a team of professional psychologists but I now understand that myself and Tim Leary were wandering around a local supermarket car park for three weeks under the impression that we were in Brazil.

Although both Leary and Ginsberg in separate phone calls have tried to convince me that I have invented it, the search for sahatasca continues.

#drugs #Alan Watts #Allen Ginsberg #Timothy Leary #oc

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instruth · 5 months ago

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An Extract Of Dementia

An extract of rich dementia

Let the bygones be bygones

Read the Book of Alzheimia’s

Step in, breathe in the Ozone

Intoxicated to the blur amnesia

Jump off the Cliff of Oblivion

Enroll in the class of extinction

Start off with forgetfulness

Revive the joy of happiness

Take a leap to the darkness

Don’t you want to remember?

Fire from snow, red from amber

Drunkenness from being sober

Falls or the White Cliffs of Dover

President from ordinary members

Sniff a doze of chloroform

Sleep on, in the dental chair

Monitored by a girl in uniform

Rub your head, count your hair

Tooth bent as worm, on golf fair

13 June 2024

A Gogyoshiren Poem (25)

Photos Credit: J. P. Album Collection

#poetryportal #writerscreeds #smittenbypoetry #spilledwords #writingthestorm #poeticstories #inkstainsandheartbeats #writtenconsiderstions

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helen0086 · 26 days ago

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Coenzyme Q10 Detection Technology

In 1957, Prof. Grane of the Institute of Enzyme Research of the University of Wisconsin isolated a new quinone compound from the lipid extract of bovine heart mitochondria [1]. The compound is an orange-yellow crystal with a melting point of 48-49 ℃, capable of reversible oxygenation and reduction, and mainly involved in mitochondrial electron transfer. The compound is coded as Q-275 (Q is the initials of quinone, and 275 is the maximum absorption at 275 nm).

In 1958, American scholar Folkers and his team synthesized a series of coenzyme Q compounds, confirmed the structure of Q-275 and named it coenzyme Q10 [2]. In 1961, Mitchell, a British chemist, proposed the theory of "chemotaxis" in the study of energy conversion in living organisms and revealed the role of coenzyme Q10 in the energy conversion system of mitochondria [3], and was awarded the Nobel Prize in Chemistry in 1978. Since then, people have gradually recognized coenzyme Q10, and its applications have been widely and deeply studied.

Coenzyme Q10 (CoQ10), also known as ubiquinone, is chemically known as 2,3-dimethoxy-5-methyl 6-deca-isopentadienylbenzoquinone and consists of a benzoquinone ring and polyisoprene side chains. The number of isoprene units in the coenzyme Q series varies by species, with humans having 10 units. Coenzyme Q10 is available in both oxidized (ubiquinone, CoQ10, Ubiquinone) and reduced (ubiquinol, CoQ10H2, Ubiquinol) forms, and its chemical structure is shown in Figure 1.

Fig. 1 Chemical structures of oxidized (a) and reduced (b) forms of coenzyme Q10

Coenzyme Q10 is an important component of the mitochondrial respiratory chain, where it acts as an electron carrier and participates in electron transfer and ATP production. Furthermore, the cellular functions of coenzyme Q10 are multifaceted: it is present in all cell membranes, it limits the toxic effects of free radicals, it is a component of low-density lipoprotein (LDL), and it is involved in the aging process. Its deficiency is associated with a variety of diseases, such as mitochondrial disease, cardiovascular disease, age-related diseases, tumors, liver disease, kidney disease, etc. Panthenol is also a powerful antioxidant. Panthenol is also a powerful antioxidant, preventing lipid peroxidation in biological membranes [4].

With the deepening of the research on coenzyme Q10, the application of coenzyme Q10 is becoming more and more extensive. In addition to its use as a drug, it also has many applications in nutraceuticals, cosmetics and dietary supplements. Coenzyme Q10 is an endogenous substance, but its concentration in living organisms is very low. The analysis and determination of CoQ10 is important for the clinical diagnosis of diseases and the quality control of drugs and health products. In recent years, many analytical methods for the determination of coenzyme Q10 have been developed, which are summarized and discussed in this paper.

1 Coenzyme Q10 extraction and sample preparation

Coenzyme Q10 is insoluble in water and methanol at room temperature, slightly soluble in ethanol, soluble in acetone, 1-propanol, and soluble in organic solvents such as hexane and chloroform. Pharmaceuticals and dietary supplements such as tablets, capsules and softgels can be dissolved in ethanol, 1-propanol and other solvents, and analyzed by ultrasonication or filtration.

The isolation and enrichment of coenzyme Q10 from complex biological matrices is a laborious process.

Conventional liquid-liquid extraction is the most commonly used extraction method. This method is simple and has a large processing capacity, but has the disadvantage of high solvent consumption and some solvents can interfere with subsequent detection. Often the solvent is evaporated under N2 protection after extraction and redissolved in a mobile phase or other solvent. Whole blood samples were immediately dosed with the anticoagulants heparin or EDTA, and the plasma was centrifuged at low temperature and stored at -80°C. The plasma was then analyzed for the presence of coenzyme Q10 in the plasma. Coenzyme Q10 was extracted from plasma as follows [5]: Methanol was added to the plasma to precipitate the proteins, and the plasma was extracted with hexane. The mixture was rotated and shaken for 15 min, then centrifuged for 5 min, and the supernatant was extracted and the solvent was evaporated. The supernatant was dissolved in acetonitrile before analysis.

Coenzyme Q10 was extracted from animal heart tissue [6]. The extraction of coenzyme Q10 from animal heart tissue [6] was performed by precise weighing, transferring to homogenization tubes containing lysis medium A (containing garnet and zirconia beads), adding 1-propanol and the antioxidant 2,6-di-tert-butyl-4-methylphenol (BHT), shaking, centrifugation, and collection of the supernatant, which was analyzed immediately. The extraction of coenzyme Q10 from muscle tissue is most often done directly using muscle homogenate, or sometimes mitochondria are extracted from the tissue under ice-cold conditions, and then the mitochondrial suspension is diluted with 1-propanol, centrifuged, and the organic layer is extracted with ethanol and hexane [7]. The one-step extraction method is to use a suitable organic solvent to extract coenzyme Q10 while precipitating proteins. Yang et al. [8] studied the one-step precipitation of plasma proteins with different organic solvents (methanol, ethanol, acetonitrile, and acetone), and found that acetone was the best precipitant, and the extraction yield ranged from 71.00% to 93.07%, and was simpler than the operation of liquid-liquid extraction.

The solid phase extraction (SPE) technique can also be used for the extraction of coenzyme Q10. On-line SPE techniques are less time-consuming, less expensive, and reduce sample loss and contamination problems. The technique is usually automated using a programmable on/off valve [9]. However, protein precipitation is required prior to extraction.

Molecularly imprinted polymers (MIPs) are specialized molecular recognition techniques that have been developed in recent years. Molecularly imprinted polymers (MIPs) are formed by mixing template molecules with functional monomers, cross-linkers and initiators. After polymerization, the template molecules are removed and binding sites and cavities complementary to the templates in size, shape and function are formed [10], allowing selective recognition and adsorption of molecules structurally similar to the templates.

Contin et al. [10] synthesized MIP using coenzyme Q0 as the template, methacrylic acid as the functional monomer, acetonitrile as the pore-forming agent, ethylene glycol dimethylacrylate as the cross-linking agent, and benzoyl peroxide as the initiator. MIP was used as an adsorbent for solid-phase extraction of coenzyme Q10 from liver samples using dispersive solid-phase extraction. In addition, MIP synthesized in the same way could be used as the filling adsorbent for solid-phase extraction of coenzyme Q10 in urine. In addition, the MIP synthesized by the same method can also be used as the filling adsorbent of polypropylene columns for solid-phase extraction of coenzyme Q10 in urine, and the columns can be reused four times [11]. Compared with the traditional solid-phase extraction, MIP as a polymer adsorbent for solid-phase extraction has the advantages of simple synthesis, low cost, good stability, porous, and high selectivity for target molecules [11].

Sometimes it is necessary to maintain the original oxygenated and reduced state of coenzyme Q10 in the samples during the extraction process, which causes great difficulties due to the oxidizability of CoQ10H2. In this case, the temperature can be controlled at a low temperature of 4 ℃ during the extraction process [6,12], shortening the extraction time and using anhydrous extract will increase the stability of CoQ10H2 [13], and the use of HCl-acidified ethanol as a diluent can also prolong the stability of CoQ10H2 and prevent the auto-oxidation of CoQ10H2 [12]. BHT is an antioxidant often added in the extraction of plasma and tissue samples, which can prevent the oxidation of CoQ10H2 [6,12,14]. However, the addition of BHT to CoQ10H2 extracts from dietary supplements and pharmaceuticals was found to increase the oxidation of CoQ10H2 [13,15]. The difference in matrix composition between plasma samples and dietary supplements may be the main reason for the loss of antioxidant capacity of BHT [13].

Biological samples for coenzyme Q10 extraction include plasma, leukocytes or platelets, muscle, fibroblasts and urine [16]. Muscle biopsy is the best choice for studying coenzyme Q10 status in mitochondrial diseases, but it is very invasive; the correlation between the levels of coenzyme Q10 and tissues in plasma, blood cells and urine has been controversial, but the determination of coenzyme Q10 in these samples has an important role in therapeutic monitoring [16]. However, the determination of coenzyme Q10 in these samples is important for therapeutic monitoring [16].

The methods used to extract coenzyme Q10 from biological samples are summarized in Table 1.

Table 1 Extraction methods of Coenzyme Q10

Simple operation, large processing capacity, high solvent consumption, some solvents may interfere with the subsequent detection.

Plasma, animal heart, muscle homogenate, mitochondria

Online Solid Phase Extraction

Less time-consuming and costly, reducing sample loss and contamination.

plasma (medicine)

Molecular Blotting Techniques

Low cost, good stability, high selectivity for target molecules, and can be combined with solid phase extraction.

Animal liver, urine

2 The main assay for Coenzyme Q10

2.1 High Performance Liquid Chromatography (HPLC)

HPLC is currently the main analytical method for analyzing coenzyme Q10 in various matrices. The main detectors coupled with HPLC are ultraviolet (UV), tandem mass spectrometry (MS/MS), electrochemistry (ECD), fluorescence (FL), chemiluminescence (CL), etc. The separation effect of HPLC is good, and each detector has its own characteristics.

2.1.1 HPLC-UV

HPLC-UV is the most commonly used method for the determination of coenzyme Q10, and has become the national standard for drugs and health foods [17, 18]. It has been widely used for the determination of coenzyme Q10 in pharmaceuticals [15, 19-22], health foods or dietary supplements [15, 20], plasma [14, 23] and tissues [10]. Conventional C18 or C8 reversed-phase chromatographic columns can separate either one form of coenzyme Q10 (usually oxidized) or both oxidized and reduced forms.

Liposomes are a new type of pharmaceutical dosage form formed by the self-assembly of lipids (mainly phospholipids and cholesterol) with a bilayer structure similar to that of a cell membrane, which can encapsulate hydrophilic or hydrophobic drugs. Ruiz-Garcia et al. [21] prepared a small monolayer of liposomes encapsulating coenzyme Q10, phosphatidylserine, and fat-soluble vitamin C (6-o-palmitoyl-L-ascorbic acid) by thin-film hydration. The prepared samples were freeze-dried, solubilized in chloroform and determined by HPLC-DAD at two analytical wavelengths.

Clementino et al. [22] prepared lecithin/chitosan nanoparticles encapsulating simvastatin and coenzyme Q10. The chitosan-modified liposomes showed higher stability and narrower particle size distribution. The content of simvastatin, simvastatin hydroxylate and coenzyme Q10 was quantified by reversed-phase HPLC-UV method to account for possible degradation products. The encapsulation rate was determined and the in vitro release of the drugs was studied. According to the study, the serious side effects of statins, such as rhabdomyolysis, were associated with the decrease of coenzyme Q10, so the co-encapsulation of these two drugs is of great significance.

Coenzyme Q10, as a fat-soluble vitamin coenzyme, is often measured in conjunction with other fat-soluble vitamins. Franke et al. [14] analyzed 25 substances including 25-OH-vitamin D3, 25-OH-vitamin D2, retinol, tocopherols, carotenoids (including their stirrup isomers), and oxidized and reduced coenzyme Q10 in plasma on a fusion-nucleated 2.6 μm particle size C18 column in tandem with a C30 column, which is good at separating carotenoid isomers, and in conjunction with a six-pass valve. D2, retinol, tocopherols, carotenoids (including their stirrup isomers), and oxidized and reduced coenzyme Q10 in plasma. The switching of the six-way valve allows coenzyme Q10 to flow from the C18 column to the detector while the carotenoid isomers are eluted on the C30 column, avoiding the difficulty of separating these two substances on the same column. In addition, if a pressure-resistant UV-Vis detector is added between the C18 and C30 columns, it is possible to separate all substances without switching the six-way valve, but special software is required to control the two detectors and to acquire and process the data. It has also been reported that retinol, six carotenoids, two tocopherols, and coenzyme Q10 (10 fat-soluble vitamins) can be measured in human plasma using a MYC30 column, and the total amount of the oxidized form of coenzyme Q10 was measured by oxidizing coenzyme Q10H2 first with FeCl3 [23].

The HPLC-UV method is highly accurate and reproducible, with LOD generally on the order of μg-mL-1 and sometimes on the order of ng-mL-1 with highly sensitive detectors [14].

2.1.2 HPLC-MS/MS: HPLC-MS/MS has been developed rapidly and applied more and more widely. This method utilizes the high separation efficiency of HPLC for complex samples combined with the high sensitivity and high selectivity of mass spectrometry, which can detect low content samples under the background of complex matrix, and is widely used in the analysis and determination of target compounds in biological samples.

The main types of tandem mass spectrometry are triple quadrupole mass spectrometry [5,7,11,25,26], quadruple linear ion trap mass spectrometry [13,24], and hybrid quadruple orbit trap mass spectrometry [12], etc. Most of them use electrospray ionization, multiple reaction monitoring (MRM), and positive ionization mode. Due to the low sensitivity of [M + H]+ analysis of coenzyme Q10, ammonium adducts, i.e., [M + NH4]+, are often used to improve the sensitivity of the mass spectrometric response. By adding a certain amount of ammonium acetate to the mobile phase, [NH4]+ forms a stable five-membered chelated ammonium cation with coenzyme Q10 [8]. The formation of Li adducts has also been reported to greatly increase the sensitivity [24].

The electrostatic field orbitrap mass spectrometry (Otbitrap) is a new type of high-resolution mass spectrometry, which has the advantages of high resolution, high mass accuracy, and wide dynamic range, etc. Pandey et al. [12] applied HPLC-hybrid quadruple orbitrap mass spectrometry (Q-Orbitrap) to rapidly determine the redox state of coenzyme Q9 and coenzyme Q10. Two scanning modes, full MS/AIF and tSIM/data-dependent secondary scanning (tSIM/ddMS/MS), were compared, and it was found that full MS/AIF had higher signal sensitivity and good peak shape. During sample preparation, coenzyme Q9 and coenzyme Q10 were extracted with BHT-containing hexane to limit the oxidation of the reduced form, and the Kinetex C18 column, with fused-core SiO2 packing and smaller particle size (2.6 μm), was found to have higher column efficiency, better resolution, and good peak shape. Oxidized and reduced forms of coenzyme Q9 and coenzyme Q10 were analyzed in brain, heart, liver, adipose tissue, and muscle of healthy mice with a small amount of sample (<5 mg) and a very short analysis time (4 min). the LOD ranged from 0.01 to 0.49 ng mL-1 .

Due to the complexity of the biological sample matrix and the low concentration of coenzyme Q10, sample pretreatment is very important. Becerra et al. [11] analyzed coenzyme Q10 in human urine by molecularly imprinted polymer solid-phase extraction (MIP-SPE) coupled with HPLC-MS/MS. The pretreatment process concentrates the coenzyme Q10 by at least 5-fold. The high degree of sample purification reduces the ion suppression caused by the matrix effect of mass spectrometry. The analytical system does not interfere with protein or white blood cell elevations, which is important in cases of coenzyme Q10 deficiency with renal impairment.

The HPLC-MS/MS method uses a lot of internal standards, and the selection of suitable internal standards is also an effective way to eliminate matrix effects. Commonly used internal standards include coenzyme Q9 [5, 11, 25], coenzyme Q4 [12], and the isotopes of coenzyme Q10, coenzyme Q10-2 H6 [7] and coenzyme Q10-2 H9 [26], which are structurally similar to coenzyme Q10. Structural analogs of coenzyme Q10, such as coenzyme Q4 and coenzyme Q9, have many advantages. They are also endogenous ubiquinones and are present in human plasma at very low concentrations, or at least at levels that do not interfere with their use in analytes at the concentrations required for analysis, and therefore do not interfere with the quantification of analytes. In addition, it separates well from coenzyme Q10 [5]. A potential source of error in mass spectrometry is ion suppression, especially in electrospray ionization mass spectrometry, where the response signal of the analyte is altered and often suppressed if an interfering substance interferes with the ionization of the analyte on the surface of the droplet, or if there is competition. The use of an isotope internal standard is a good solution to the problem of ion suppression. By co-eluting the isotope internal standard with the analyte, the effects of various effects can cancel each other out, and the matrix effect can be minimized and the sample recovery can be better [7].

2.1.3 HPLC-ECD

Electrochemical detectors (ECDs) are widely used because of their high sensitivity, good selectivity and low price. Coenzyme Q10 can undergo a reversible redox reaction and can be detected by an ECD.

The commonly used detection methods are coulometric or voltammetric analysis. Different voltages are set according to the redox potentials of the substances to be measured. For oxidized coenzyme Q10, it is usually reduced to its reduced form first, and then oxidized as the original reduced coenzyme Q10 in the sample. This method can measure both oxidized and reduced coenzyme Q10 simultaneously.

Yubero et al. [27] used HPLC-ECD to determine coenzyme Q10 in urine and gave reference values for the pediatric population. An ESA Coulochem II electrochemical detector was used, and the cell voltages were -600 mV and +600 mV. The amount of coenzyme Q10 in urine fluctuated greatly at different times of the day, and the morning urine with the smallest fluctuation was chosen as the sample. The results were expressed as the amount of coenzyme Q10 per gram of particulate protein. The reference standards for children are: 2-10 years old: 24-109 nmol; 11-17 years old: 43-139 nmol. This assay provides a noninvasive method for assessing renal coenzyme Q10 status in patients with renal disease, but it is not currently available and requires up to 30 mL of urine per sample.

Schou-Pedersen et al. [6] determined reduced and oxidized coenzyme Q10 in canine plasma and cardiac tissue by HPLC-ECD and compared it with HPLC-MS/MS. The ECD was performed by fluid dynamic voltammetry using an RS6011 ultra-analytical cell at a voltage setting of 500 mV. A guard cell at -600 mV was used prior to the analytical cell to reduce oxidized coenzyme Q10 eluting from the column. Mass spectrometry was performed using a Waters Micromass Quattro micro API triple quadrupole mass spectrometer with multiple reaction monitoring (MRM) and the internal standard CoQ10-2 H9. Both methods used the same column with slightly different mobile phase ratios and additives. The results showed that CoQ10H2 was approximately 30% lower in the HPLC-MS/MS method than in the HPLC-ECD method, which may be due to differences in the calibration stock solutions or to accelerated oxidation during storage or analysis in the LC-MS/MS system. Therefore, the two methods are not interchangeable. In terms of sensitivity, the sensitivity of the two methods was comparable for coenzyme Q10H2, whereas the sensitivity of the HPLC-ECD method was higher for coenzyme Q10.

2.1.4 HPLC-FL and HPLC-CL

HPLC with a fluorescence (FL) detector is widely used for the determination of various substances in biological samples due to its high selectivity and sensitivity. Coenzyme Q10 is not a fluorescent substance and needs to be derivatized before determination. Nohara et al. [28] measured CoQ10 and CoQ10H2 in blood by post-column derivatization using HPLC using 2-cyanoacetamide and CoQ10 and CoQ10H2 heated under alkaline conditions to produce fluorescent products. The fluorescence emission and excitation wavelengths were 442 nm and 549 nm, respectively.

HPLC coupled with a chemiluminescence (CL) detector has also been reported for the determination of coenzyme Q10.Kishikawa et al. [29] used dithiothreitol (DTT) as a reductant to reduce quinone to semiquinone radicals, and semiquinone radicals converted dissolved oxygen to superoxide anion, which reacted with luminal to form CL.Accordingly, coenzyme Q10 was determined in plasma by HPLC-CL, and other components in plasma were not interfered with. Coenzyme Q10 in plasma was determined by HPLC-CL, and other components of plasma were not interfered.

Both methods require a reaction coil between the column and the detector, and require two or three pumps to mix the various reaction reagents with the coenzyme Q10-containing eluent after the column and then into the reaction coil, which is a cumbersome operation. In recent years, the literature in this area is relatively scarce.

2.2 Spectrophotometric and fluorescent methods

The Enzyme Labeler, also known as Microplate Reader, is an instrument for reading and analyzing the results of Enzyme Linked Immunosorbent Assay (ELISA) experiments. The basic principle of ELISA is similar to that of spectrophotometer or photoelectric colorimeter, using plastic microplates instead of cuvettes, usually 48-well, 96-well, or larger, with low reagent consumption, high speed, and good repeatability. Multifunctional enzyme labeling instrument often has a variety of detection functions such as absorbance, fluorescence, chemiluminescence, etc., in the medical and health inspection has been widely used.

Fukuda et al. [30] developed a rapid microtiter plate method for the determination of coenzyme Q10 using the redox cycle of quinone. Coenzyme Q10 was reduced to ubiquinone radical by NaBH4, and then the ubiquinone radical was oxidized to ubiquinone and superoxide anion radical by dissolved oxygen. The superoxide anion radical converts 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl-2H-tetrazolium chloride (INT) into a pink methanol dye. It has a strong absorbance at 510 nm, which increases with increasing concentrations of coenzyme Q10. The absorbance of Mazanine dye was measured quickly and easily by a microplate reader. As an application of this method, the content of coenzyme Q10 in cosmetics was successfully determined with an LOD of 14.8 nmol-L-1 . The proposed method can be used for the rapid high-throughput analysis of ubiquinone-containing products.

Fei et al. [31] developed a new method for the determination of coenzyme Q10 in serum and urine of Alzheimer's disease patients by fluorescence spectrophotometry, also using a microplate reader. The method is based on the fact that the chemical derivative between ethyl cyanoacetate (ECA) and coenzyme Q10 is fluorescent and can be detected by fluorescence spectrophotometer (FS-ECA) at λex/em = 450/515 nm. The results showed that serum and urine levels of coenzyme Q10 were significantly lower in Alzheimer's patients than in age-matched controls. This method has similar LOD and LOQ as the HPLC-UV method.

The FS-ECA method has some advantages over the HPLC-UV method, such as easier sample preparation, faster detection speed, and similar accuracy and specificity [31].

The important role of liposomes as a new drug dosage form for co-administration and targeted delivery was described in the literature [21,22], and liposomes can also be used as micro-containers to protect and concentrate reagents.Román-Pizarro et al. [32] prepared a new type of magnetic liposomes (MLs) containing hydrophobic magnetic gold nanoparticles (Fe3 O4 @ AuNPs) and the long-wavelength fluorophore cresyl violet for the determination of coenzyme Q10 in foodstuffs. AuNPs) and a long-wavelength fluorophore, cresyl violet, were used for the determination of coenzyme Q10 in food. First, the MLs were introduced into the flow-through system using a flow injection device and retained in front of the detector for 300 s by means of a solenoid device to achieve preconcentration. Then, a coenzyme Q10 solution containing the surfactant Triton X-100 was injected into the flow-through system. The surfactant caused the solubilization of the MLs and the release of cresyl violet, which was oxidized by coenzyme Q10, resulting in a decrease in the fluorescence signal. The concentration of coenzyme Q10 is directly proportional to the decrease in fluorescence signal. The LOD of this method is lower than that of the LC-UV method, but the equipment required is simpler and less expensive.

2.3 Electrochemical analysis

The redox properties of CoQ10/CoQ10H2 allow the determination of coenzyme Q10 by electrochemical analysis. The methods are generally voltammetric, such as cyclic voltammetry (CV) [33], differential pulse voltammetry (DPV) [34], square wave voltammetry (SWV) [35], etc. The samples can be pharmaceuticals, dietary supplements, animal and plant tissues, etc. The samples can also be used for the determination of CoQ10/CoQ10H2. Samples can be pharmaceuticals, dietary supplements, plant and animal tissues, etc.

Li et al. [34] investigated the electrochemical reduction mechanism of coenzyme Q10 at a silver electrode and developed a DPV method for the direct determination of coenzyme Q10 in plant and animal tissues. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) revealed that the reduction of coenzyme Q10 under anoxic conditions is a reversible one-electron, one-proton reduction and forms a stable semiquinone radical (coenzyme Q10H-), which is quenched by oxygen in an oxygen-filled environment. This is the reason why coenzyme Q10H2 is able to scavenge oxygen radicals due to its antioxidant function. Under the optimized experimental conditions, the DPV method can be used to determine coenzyme Q10 in complex samples, and it is rapid, sensitive, and highly selective, with an LOD of 3.33 × 10 -8 mol-L-1 .

Graphene, a single atom thick planar sheet composed of carbon atoms heterogeneously linked by sp2 in a honeycomb lattice, is a new type of sensor material [35]. Screen printing is a practical technique for manufacturing low-cost disposable sensors [35].

The new graphene sensor developed by this technology can be used for the determination of coenzyme Q10 and α-lipoic acid. The MnO2-modified screen-printed graphene electrode (MnO2/SPGE) has a larger capacitance and electrically active surface area, which facilitates electron transfer and significantly improves the oxidation performance of coenzyme Q10 and α-lipoic acid. The MnO2-modified screen-printed graphene electrode coupled with square wave dissolution voltammetry (SWV) can be used for the simultaneous determination of coenzyme Q10 and α-lipoic acid in dietary supplements with high sensitivity and practicality.

The electrochemical mechanism of coenzyme Q10 is complicated by the different electrodes and media. In a 1.1:1 methanol-ethanol solution, the electrochemical reaction of coenzyme Q10 at the glassy carbon electrode was controlled by adsorption, and the sensitivity of the determination could be improved by pre-enrichment [33]. In anaerobic ethanol solution, the cathodic process of coenzyme Q10 at the silver electrode was one-electron-single proton reduction [34], while the oxidation on MnO2/SPGE showed two-electron-single proton transfer [35]. Michalkiewicz [36] investigated the anodic oxidation of oxidized coenzyme Q10 in acetic acid solution using a glassy-carbon electrode and a carbon-fiber microelectrode coupled with voltammetry, respectively. The oxidized coenzyme Q10 in acetic acid solution was studied by

The results show clear oxidation peaks or waves in the potential range above 1.5 V. The presence of these signals cannot be linked to the well-known redox pair CoQ10/CoQ10H2, but may be attributed to the irreversible and diffusion-controlled two-electron oxidation of methoxy in coenzyme Q10 (formation of two additional quinone groups at the 2 and 3 positions of the ring). the total number of electrons involved in the CoQ10 anodic oxidation is much greater than two, suggesting that the oxidation also takes place in the unsaturated isopentadienyl side chain. The total number of electrons involved in CoQ10 anodic oxidation is much higher than two, suggesting that oxidation also occurs in the unsaturated isoprene side chain. The oxidation of the oxidized coenzyme CoQ10 has been rarely reported, is much less readily accessible than that of coenzyme Q10H2, and the mechanism of oxidation has yet to be demonstrated.

2.4 Other analytical methods

Supercritical fluids are substances whose temperature and pressure exceed the critical point, in a state of gas-liquid indistinguishability, with a density close to that of liquids, and a viscosity close to that of gases, and a high diffusion coefficient. Supercritical fluids with high diffusivity and low viscosity, very suitable for mobile phase. The supercritical fluid with more research and application is supercritical CO2. Supercritical fluid chromatography-tandem mass spectrometry (SFC-Ms/Ms) with supercritical CO2 as mobile phase can be used for the determination of coenzyme CoQ10 in rat plasma [8]. The method uses one-step acetone method to precipitate the protein and extract CoQ10 from the sample. Due to the low sensitivity of [M + H]+ of CoQ10 in mass spectrometry, methanol containing ammonium acetate was used as a post-column compensating solvent to provide [M + NH4]+ and improve the sensitivity. Supercritical CO2 is non-toxic, non-flammable, and relatively inexpensive, so it is widely used. Due to its non-polar nature, it is well suited for the analysis of fat-soluble compounds and can greatly reduce the use of organic solvents.

Other analytical methods include: high performance thin layer chromatography (HPTLC) [37], Fourier transform near infrared spectroscopy (FT-NIR) [38], nuclear magnetic resonance hydrogen spectroscopy (1 H- NMR) [39], etc. HPTLC is simple and rapid, but the sensitivity is not very high, and can be used for the analysis of coenzyme Q10 in raw materials and pharmaceutical preparations. FT-NIR does not require complex sample pretreatment, but requires a certain number of samples to establish a calibration model and obtain results through complex statistical analysis, and is generally used for the initial screening of the target analyte. 1 H- NMR also does not require complex sample pretreatment, can be both calibrated models, and obtained through complex statistical analysis, and is generally used for the initial screening of target analytes. FT-NIR does not require complex sample pretreatment, but requires a certain number of samples to establish a calibration model and obtain the results through complex statistical analysis, and is generally used for the initial screening of target analytes.1 H-NMR also does not require complex sample pretreatment, and can be used both qualitatively and quantitatively, with low sensitivity, and can be used for routine analysis of preparations. These methods can be used as a useful supplement to the quantitative analysis of coenzyme Q10.

3 Simultaneous determination of oxidized and reduced coenzyme Q10

In many methods, the total amount of coenzyme Q10 is determined by adding oxidizing agents such as FeCl3 to oxidize coenzyme Q10H2 to the oxidized form, and then the total amount of the oxidized form is determined. However, coenzyme Q10 coexists in both oxidized and reduced forms in biological matrices, and sometimes it is necessary to determine the two forms of coenzyme Q10 in biological samples, drugs and supplements separately. Commonly used methods include HPLC-UV [14, 15], HPLC-MS/MS [12, 13, 24, 25, 40], HPLC-ECD [6, 41], HPLC-FL [28], and so on.

Coenzyme Q10H2 standards are sometimes not readily available and can be obtained by reducing oxidized coenzyme Q10 with reducing agents such as NaBH4 [12-14,24,25,28,40,41]. The reduction of coenzyme Q10 at low concentrations may be incomplete, even if the amount of NaBH4 is 8800-fold excess [13], so the reduction process needs to be controlled at a certain concentration range. The reaction is usually carried out at low temperature and in the dark, and sometimes ED-TA is added to the solution after the reaction [12,13,15,25,41], which mainly binds to the metal ions that catalyze the oxidation process and acts as an antioxidant. Even if a standard of coenzyme Q10H2 is available, it may be partially oxidized and needs to be reduced before use [15], or the absorbance of the stock solution (ε = 4010) can be measured spectrophotometrically at 290 nm to determine the exact concentration [6].

Whether the quantitative results were expressed as the total amount of coenzyme Q10 or as the oxidized and reduced amounts, respectively, could affect the sample preparation. In order to maintain the reduced state of coenzyme Q10H2, in addition to the rapid operation at low temperature and the addition of antioxidants such as BHT during the preparation process, researchers have different opinions on whether the extracted solution should be re-dissolved by evaporating the solvent in the presence of N2. Some of them evaporate the solvent and re-dissolve it during sample pretreatment [12,14,15,24], while some scholars believe that there will be significant oxygenation of Coenzyme Q10H2 during solvent evaporation, so the extracted solution should be immediately dissolved in the presence of N2 [12,14,15,24].

Determination [6 ,13 ,25 ,28 ,40 ,41].

In fact, coenzyme Q10H2 is highly unstable during extraction and determination.Yamashita et al.[41] found that coenzyme Q10H2 was stable only at -78 °C, and the rate of oxidation of coenzyme Q10H2 to coenzyme Q10 increased with increasing storage time and temperature.Claessens et al.[25] found that significant oxidation of coenzyme Q10H2 to coenzyme Q10 occurred after 2 h in 1-propanol extracts of human plasma, so routine analysis was limited to 12 samples per batch in order to keep the total run time within 2 h. In addition, coenzyme Q10H2 is not stable at the same temperature as coenzyme Q1010. Claessens et al. [25] found that in 1-propanol extracts of human plasma, significant oxidation of coenzyme Q10H2 to coenzyme Q10 occurred after 2 h. Therefore, routine analyses were limited to 12 samples per batch in order to keep the total run time within 2 h. The results of this study are summarized below.

Due to the uncontrolled nature of oxidation, it has been suggested that almost all mitochondrial coenzyme Q10H2 is oxidized during sample pretreatment, and therefore quantification of total coenzyme Q10 in isolated mitochondria does not require ubiquinol oxidation [7]. Nevertheless, attempts have been made to control the rate of oxidation of coenzyme Q10H2 during analysis or to know the extent of oxidation. The choice of internal standards has helped to realize this desire. Structural analogs of coenzyme Q10 and coenzyme Q10H2, such as diethyl- or dibutyl-coenzyme Q10 [14] and dipropoxy-coenzyme Q10 [40], are sometimes used as internal standards in the analysis of coenzyme Q10 and coenzyme Q10H2. They are structurally very similar to CoQ10 and CoQ10H2 and exhibit the same chemical behavior as the analytes, especially with respect to artificial oxidation, which makes it possible to accurately back-calculate the original CoQ10H2/CoQ10 ratio [14].

The CoQ10H2/CoQ10 ratios in biological tissues varied, and Claessens et al. [25] showed that the plasma CoQ10H2/CoQ10 ratios in healthy volunteers without nutrient supplementation ranged from 22.3 to 64.4, with an average of 41.7, whereas Yamashita et al. [41] showed that the plasma CoQ10H2/CoQ10 ratio was about 95/5, suggesting that plasma coenzyme Q10 is mainly in the reduced form. These results indicate that plasma coenzyme Q10 exists mainly in the reduced form.

Changes in the CoQ10H2/CoQ10 ratio have important physiological significance and are associated with many functional disorders and diseases. Measurement of the CoQ10H2/CoQ10 ratio is useful in exploring the mechanisms of many diseases. Oxidative stress has been defined as a disturbance of the pro-oxidant-antioxidant balance in favor of the former, and is considered to be a causative factor in aging and degenerative diseases such as cardiac diseases, diabetes mellitus and cancer [41]. There is a consensus that the CoQ10H2/CoQ10 ratio is an important parameter in the assessment of oxygenation stress [24, 25, 41].

Another study showed that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TC-DD) damaged mouse liver in a dose-dependent manner. Tang et al. [40] investigated the mechanism, and found that exposure of mouse liver samples to TCDD resulted in a decrease in the total amount of coenzyme Q10, a decrease in the level of coenzyme Q10H2, and an increase in the CoQ10H2/total CoQ10 ratio. This may be due to the inhibition of succinate dehydrogenase in the electron transport chain. In addition, the decrease in the total amount of CoQ10 implies that CoQ10 is degraded by external environmental influences, which was confirmed by Temova Rakua et al [42]. They found that oxidized coenzyme Q10 was degraded during storage of dietary supplements and drugs containing coenzyme Q10, and that oxidized coenzyme Q10 was converted to reduced coenzyme Q10H2, especially in the presence of antioxidants such as vitamin C.

4 Summary

Coenzyme Q10 is an important electron carrier and antioxidant component of the mitochondrial respiratory chain and is widely found in human cells. Coenzyme Q10 deficiency may be associated with a variety of diseases. Although it is an endogenous substance, it can be used as a drug or dietary supplement to treat or ameliorate certain related diseases. Therefore, the selection of efficient isolation and analytical methods is of physiological and clinical importance. Liquid-liquid extraction is the most common method for the extraction of coenzyme Q10, while the extraction of reduced coenzyme Q10H2 requires temperature control and the addition of antioxidants. Solid-phase extraction and molecular blotting techniques have also been applied in the extraction of coenzyme Q10 from biological samples, which have greatly improved the extraction efficiency and detection sensitivity.

Coenzyme Q10 can be detected by a variety of methods, and currently the most commonly used method is HPLC. In clinical and pharmaceutical analysis, miniaturization of instrumentation by reducing column diameter and length and particle size is one of the major trends in improving separations [43]. HPLC-UV is easy to use as a standard method, has good stability, is not very sensitive, but is generally sensitive enough to meet the requirements for the simultaneous analysis of a variety of components.

HPLC-MS/MS has high sensitivity and good selectivity, and has unique advantages for the analysis of coenzyme Q10 in complex matrices, such as biological samples, but the operation of the instrument is complicated and the price is expensive. The electrochemical analysis method is simple, fast and sensitive, and has certain applications in the analysis of coenzyme Q10. The HPLC-ECD method is convenient for the simultaneous determination of oxidized and reduced Coenzyme Q10. Coenzyme Q10 co-exists in both oxidized and reduced forms in almost any sample. Some analytical methods are capable of determining both the total amount of coenzyme Q10 and the oxidized and reduced forms, while others can only determine the total amount, depending on the HPLC separation. The characteristics of the various methods, their determination formats and their applications in samples are shown in Table 2.

Table 2 Comparison of different analytical methods for Coenzyme Q10

Easy and fast to use, sometimes requires color development or derivatization, matrix may be interfering

Coenzyme Q10 is mainly present in reduced form in organisms, and the ratio of CoQ10H2/CoQ10 is clinically important, with greater bioavailability of CoQ10H2 in drugs and dietary supplements. Therefore, the simultaneous determination of oxidized and reduced coenzyme Q10 is a future development. The distribution of the two forms, their interconversion and their biological significance will be a focus of future research, which also brings opportunities and challenges to the study of extraction and analytical methods for both forms of coenzyme Q10.

In order to meet the clinical needs, coenzyme Q10 can be prepared by microbial fermentation [44] or chemical synthesis in addition to extraction from biological samples. Chemical synthesis is divided into total synthesis [45] and semi-synthesis [46], and the intermediate of semi-synthesis is ganiol. Microbial fermentation can be used for large-scale industrial production.

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