Autophagy

our collaborator has absolutely LOST IT at their latest manuscript submission’s reviewer comments. this is his response back to the editor. like dAMN. especially that last sentence?? lol get REKT.

text (with my bold for emphasis on the juiciest bits):

Dear Dr. [    ],

I want to express my thanks to you for overseeing the review process of our manuscript. Unfortunately, I find the reviewer comments of little help for our attempts to improve the manuscript. The reviewers did not provide a constructive or comprehensive critique of the specific approaches and results presented in our study. All three critiques used generic boilerplate and standard comments, to be summarized by the notion that we did not perform all the experiments that the reviewers were able to think of.

Almost any published paper has remaining open issues that need to be addressed. I would even argue that it is a key feature of a good paper that it opens up new questions rather than closing a case. In our specific case, we provide a mechanistic analysis of how curcumin isomers affect the NF-kB pathway, and by using our unique approach, we have identified a key difference in the effect of curcumin versus its isomer BDMC. This difference has never been worked out from a mechanistic perspective which is exactly what our manuscript has accomplished. From that point of view, in the revised manuscript, we are not going to perform additional studies in animals, or explore effects on autophagy (why?), or analyze nuclear translocation (not necessary since we are using a functional assay anyway). 

This just to argue against some of the reviewer comments. In addition, a response to all of the reviewers comments is provided below. 

By focusing on mechanistic rather than phenomenologic analyses, our work has been unique in the field and of consistent high quality, both in its science and writing. I have published a number of papers in ACS journals, and I don’t think this manuscript will be the kind of disgrace to your journal as has been invoked by the referees.

Kind regards,
[     ]

Orochimaru One Shot: autophagy

Happy >Birthday, baby <3 - 10/27/2017

“Damn!”

The clash from a broken syringe echoed through the room. The injected skin of the boy turned purple and reminded vigorously of poison. The pale, bound arms tried to struggle, the chewed off fingertips dug into the metal table while the involuntary patient tried to let out a scream which ended in a hoarse cry.

Orochimaru let out an annoyed sigh and allowed his tired eyes to rest for a second while Kabuto came running to silence the boy. The expectations had been very high this time. He had thought that this would be the cure, the end of sleepless nights and weeks he had spent in the lab hissing at his subordinates. Rustling up a live organism with the same disease had been difficult enough.

He needed to get away from this failure. Leaving the struggling boy behind, he pushed the heavy doors open and left the lab. The dying light that shone through two bullseye-shaped windows illuminated the back of his head and refrained this way from stinging his eyes while the troubled man moved forwards. The dark creepy hallways seemed harmless next to the snake-like scientist.

He moved past the prisoners who hurriedly scurried into the back of their cells as soon as they saw the black haired man walk by. Orochimaru couldn´t care less if they feared him. Fear meant respect, recognition. The distance to his destination grew smaller and smaller. Which each step he shed a piece of his grudge because of the incident in the lab.

As soon as the heavy door opened, Orochimaru´s shoulders slacked off as he felt the stress melt away from his mind and body. His gaze was already focused on the young woman stirring in the sheets, his sharp pupils dilating. It seemed like the girl roused from a half-sleep full of tumultuous dreams. She let out a whine that was muffled by a face mask and her movements were constricted by a straitjacket. Orochimaru approached her and sat down next to her on the bed.

“Good morning~.”

He was completely changed. His voice spoke in a warm, soft tone. He released her from the restrains and gathered her up in his arms with a soft glance. As he did, the fabric uncovered the bite marks and gnawed off skin of her fingers and arms. The girl protested silently and rubbed her eyes with cramped movements. Her arms and hands tingled from being held in one place for so long and she started to sob.

“Shhh, darling.” Orochimaru cooed while he rocked the girl in his arms. She continued to sob which he hushed with a quick kiss to her badly bitten lips. The young woman squirmed a little but relaxed after a few moments, sinking into his hug. Whether it was from tiredness or affection didn´t matter.

“Good girl.” Orochimaru let a smile pull at his lips while he held her tight and ravished in her presence. With her next to him he felt at ease. She was his precious little secret.

His long fingers combed through her hair while his eyes darted over her features. Did her condition improve? Aggravate? Did the side effects of her countless medication start to show? So many things he had to worry about. His nails peeled some caked blood away from her chin. He despised dirt.

“I will order someone to draw you a hot bath. Would that be good?”

While Orochimaru held the young woman, her regular breathing slowed down until she seemed to nap against his shoulder.

“Yes, go to sleep. I´ll be visiting you again tomorrow.”

And with that, he put the ties back on and kissed her goodbye, letting sleep take her back into its safe arms. He felt just as bound as her as he walked back into the lab to experiment for what would be another sleepless night.

She was his dark little secret, waiting for him in the most hidden place of his hideout. With every day her disease affected her brain further, at this point she would try to eat up her fingers and hands if no one kept her restrained. Even so, he would do everything in his might to keep his precious girl alive and well.

The weeks rolled by without any further improvement. Just when the formula was almost finished completely, they were faced with bitter setbacks that slowed down completion.

Another failed attempt. Orochimaru wiped the sweat off his forehead. The bright light of the neon tube blinded him as he leaned his head back to take a deep breath. He needed to see her. Now.

The heavy door opened with a creak and Orochimaru rushed in.

“Good morning, my dear.”

The familiar rustling of sheets failed to appear. A little worried, the tall man sat down on the bedside and pulled down the sheets to reveal the young woman sleeping.

“Wake up, darling, it´s morning.” His amused voice hid the worry that rose with every second.

She didn´t open her eyes, only scrunched her eyebrows together instead. Orochimaru pulled the blanket down further and took off her straitjacket as well as her face mask in a hurry and scooped her up in his arms. She still continued to sleep as he brushed the side of her face.

“Are you not happy to see me?” Something in his voice broke, while his eyes scanned her body to find a reason for this unusual behaviour. He unconsciously held her tighter, waiting for her to open her eyes, which she didn´t.

He felt strange. His heart beat like the one of a scared bird while his eyes darted over her fragile body. It had to be the medicine. In order to keep her safe in this room, the drugs helped her stay asleep most of the day. Usually it wasn´t a problem to wake up when Orochimaru visited her, but right now that seemed to be the case.

The medicine had been Kabuto´s idea. Orochimaru´s eyes narrowed in anger as he set the young woman back on the bed and put the blanket on her.

In a minute he was back in the lab. A metal table flipped over and in the next second, Kabuto was pinned to the wall by his master that had grabbed him by the collar.

“You said the drugs wouldn´t hurt her.”

Instead of fear, a smirk tugged at the assistant´s lips. “I´m sorry, my lord. I tested the medicine on several patients before and I can assure you that it worked perfectly.”

The grip around his throat tightened as sharp yellow eyes stared daggers into a pair of glasses. He had never talked to Kabuto like this. Her condition had to really get to him.

“Do something.” It came out in a low hiss, fangs baring while snake-like eyes shone with threat. After these words, Orochimaru released the hold on his assistant and walked into his quarters to get some sleep.

After two weeks, they finally found the formula. It was already too late. As Orochimaru removed the seals to get into the room, a strange apprehension let him freeze to the spot.

Her head peeked out from under the sheets. Her body didn´t seem to move and also her chest didn´t rise up nor down. Orochimaru slowly walked to her bed and sat down, holding her in his arms for what would be the last time. He had to blink to see clearly, stroking over her head for what felt like hours. The wet feeling on his cheeks was new. She was dead. And the last bit of his humanity had died with her.

After the black haired man left the room, he returned to his lab. Kabuto looked up from some paperwork, eyes fixed on his master´s expression. Why was he back so soon?

“Get someone to take care of her body.”

The words didn´t fail to bring a smirk onto the assistant´s lips. As he was about to walk out of the lab, when his master´s voice stopped him.

“Kabuto. Close the files about the whole project. We will focus on our main goal again.”

Kabuto nodded, seemingly pleased with the orders. He opened the door and stepped out, being just about to close the door.

“That was what you wanted, wasn´t it?”

Orochimaru´s words had him freeze to the spot at once. The fear vanished from him after a few seconds however. If Orochimaru wanted to kill him, he would already be dead. Just how had he found out about this?

The black haired man watched his assistant walk out of the door and turned to the messy desk. Soon there wouldn´t be any trace left of the work they had accomplished down here. It would be like the woman never existed in the first place. Kabuto must´ve thought his master was wasting too much time on this pointless project. It was quite amusing how far his assistant went to direct Orochimaru to the right channels, even though his raspy laughter got betrayed by just the smallest sting in his heart.

What Happens to You During a 3 Day (72 hour) Fast/ How to Fast for 3 Days

This is going to be a lengthy, informative post so if you’re looking for something else, keep scrolling but I would highly advise you read it if you plan on fasting! Disclaimer: I am NOT pro anything, but I cannot forcibly stop people from doing things, all I aim to do is inform people so they don’t get hurt. If you have an issue with me informing others so they stay safe even while having an ed, please block me. Now for the real post!~

The first thing you need to know is this: a 3 Day (72 Hour) Fast is safe, healthy, and effective. If you are underweight/malnourished, this changes of course but for those looking to lose a few pounds, a three day fast could be the thing for you.


When tapped into a physiologically fasted state, your body goes through a series of adaptations.


#1: Cellular Detoxification and Repair

While feeding, our bodies don’t have the desire nor the need to thoroughly cleanse itself. The main mechanism for this is autophagy (a metabolic pathway that causes the organism to “self-digest” and removes waste material from the cells.) This is required in order to maintain lean body mass and it prevents the breakdown of muscle in adults. It is also needed for healthy brain cell mitochondria. Inflammation and stress are reduced as well, which helps fight off illnesses. Increased levels of glucose (a simple sugar that is a component of many carbohydrates), insulin (a hormone produced in the pancreas that regulates the amount of glucose in the blood. Lack of insulin causes a form of diabetes), and proteins all turn off autophagy (even just 3mg of a raise). To release autophagy, your liver glycogen (a substance deposited in bodily tissues as a store of carbohydrates) stores need to be depleted first. This takes up to 14-20 hours of fasting and will only reach its peak after 48 hours.

#2: Boosts Growth Hormone and Retains Muscle

While fasting, the body shifts into a state of nutritional ketosis (a metabolic state involving the production and use of fatty acids and ketone bodies) which torches fat because you’ll be drawing all your energy from the adipose tissue (tissue used to store fat). Extended fasts aren’t as catabolic (the breakdown of complex molecules in living organisms to form simpler ones, together with the release of energy; destructive metabolism) as you might think. The biggest fear people have in regards to fasting is that you’ll lose muscle. This won’t happen because:

  • Ketosis is anti-catabolic and decreases a need for glucose because it makes you gain energy from fat instead.
  • Fasting triggers Human Growth Hormone (HGH) which is responsible for building, maintaining and preserving lean tissue (including muscle, bone, ligaments, tendons, and other organs)

HGH increases within the first 16+ hours of fasting by 1300-2000%. Only after incredibly extensive fasts (10+ days) will you see HGH flatten back out. Moral of the story is a 3-day fast = 0 muscle loss!

#3: Increased Metabolism

Contrary to some beliefs I’ve seen floating around online, Intermittent Fasting (a diet that focuses on having defined periods of fasting and a period of eating) does increase metabolism by roughly 3.6% after 48 hours! A 4-day fast would see an increase of 14%. Fasting increases your metabolism and puts it into temporary “high gear”. You may, however, experience an initial slump in your metabolism because your body becomes more conservative with its fuel.

#4: Better Biomarkers

 Insulin is a key hormone in regards to storage and distribution of nutrients in the body. It is released by the pancreas in reply to food intake. When blood sugar rises, insulin is released and lets your cells take that sugar in to store energy and prevent elevated blood sugar levels. Most heart diseases are caused by high blood sugar and insulin levels. You want insulin to be lower and fasting is the best way to achieve that. Fasting can help fight diabetes, lower blood sugar levels, improve insulin sensitivity/effectiveness, reduce cholesterol as you aren’t taking in any carbs, reverse insulin resistance (insulin resistance leaves blood sugar levels high-it’s bad), decreases synthesis of triglycerides (derived from glucose and fatty acids, contributes to body fat), promote stem cell growth which can increase longevity (stem cells regenerate and repair damaged tissue), and it may lengthen your lifespan, too!

#5: Protection Against Cancer and Tumors…

Autophagy can purge precancerous cells and cause the cellular death of malignant tumors. This is triggered by fasting for a long time. Dr. Dominic D’Agostino, a leading expert in cancer research, fasting, and nutritional ketosis advises everyone to have a fast of 3-5 days, 2-3 times per year (provided you aren’t underweight or anything bad-just ask your regular doc to be safe). When you fast, you’re conducting deep cellular cleansing that reduces the risk for cancer by a large margin!

#6: …And Neurodegenerative Disease

Fasting ups levels of ‘brain-derived neurotic factor’ (BDNF) hormone, which a deficiency of is implicated in depression and similar issues. When BDNF levels are up, new brain neurons are made- called neurogenesis. Neuroplasticity (the ability to firm neural connections) benefits from this too. Fasting supports brain power and gives protection for neurodegenerative disease while mitigating cognitive decline (translation: it makes the chance of diseases like Parkinson’s or Alzheimer’s much less severe because it makes new brain nerve cells and helps boost your brain power)

#7: Lengthened Lifespan???

Fasting can improve longevity. If autophagy and HGH are rampant, your body will experience less stress, inflammation and muscle loss which are the biggest factors in aging. Mice’s lifespans were extended however no human studies have been conducted. If your body experiences less stress, it can build a better response to it and thus cope with it better. When you don’t give an organ calories, the lifespan increases by 30%, but you would, of course, eventually starve to death. The key is to do it intermittently!

3 Day Fast Without Getting Hungry?!

#1: The First 24 Hours: Have a low carb meal the night before so your glycogen levels are depleted already, thus entering ketosis sooner and inducing autophagy that much quicker. Drink a cup of water with a tiny pinch of salt when you wake up to replenish electrolytes. Wait 1-2 hours and then drink a cup of green or herbal tea. For black tea, wait 2-4 hours because of the caffeine. Do NOT add artificial sweetener!! Drink sparkling water to reduce hunger. When you feel hungry for the first time, drink more water and wait 15mins. If the hunger persists, drink a cup of black coffee, You can have decaf throughout the day, but never as a substitute for water or tea. Your max coffee intake while fasting is 2-4 cups due to the risk of dehydration. Avoid diet sodas while fasting or any other artificial sweetener as they raise insulin levels. Be productive, go for a walk, etc. When/If the hunger gets worse, use Apple Cider Vinegar (if you hate the liquid’s taste, Walmart has a big bottle of ACV pills for like 6$) and brush your teeth (alternatively, drink peppermint tea, the mint suppresses appetite). When you reach the 20 Hour mark, drink a warm glass of lemon water. You can choose to add in 1/3 teaspoon each of turmeric, ginger, and cinnamon to add flavor (they also all have some pretty good health benefits to look into but I won’t be listing them since this is already a hella big post). This lemon water will act as a “dinner” of sorts. The first 24 hours of your fast are usually the hardest if you’re used to eating a normal or near-normal amount of calories (personally, I found that near 48 hours was the hardest for me, and I usually restrict under 1000cals per day, most of the time around 500cals per day) Your body is still geared toward burning glycogen stores until they’re depleted and your body enters ketosis. 

#2: 24-48 Hours, The Second Day: If you make it to bed without feeling really bad hunger and pain, congrats you can consider your fast a success already. It’s all downhill now because your body is buzzing with ketones. Follow the same steps as Day 1 (water, avoid diet sodas, etc.). Your mind should still be sharp and clear at the 30-hour point. (Personally, it wasn’t until 40 hours in that I started feeling really tired and like I couldn’t focus on just one thing at a time- my head was everywhere and nowhere all at once but I also have ADHD, so maybe that’s just me!) There are some limits, but given that you haven’t eaten for a full day, you should feel pretty damn great. The elevated ketones lead to acetonic acid that produces a mild intoxication similar to ethanol. What gets people most during fasting are social pressures and food signals. You get the urge to quit and just give in but keep going! Your insulin levels can increase just looking at food (that darn placebo effect!) So unless you can sit in front of a plate of food with absolutely no desire to eat it, avoid environments with that kind of stimulation. (If you really want to you can look up how to do that online, there are plenty of other anas who have shared that info- to get better results on searches you can put ‘pro-ana’ in front of it just take what they say with a grain of salt, kids.)

#3: 48-72 Hours, The Final Day: Going to bed on the second day is usually easier than on the first (I didn’t get to do this due to feeling dizzy and my family and gf becoming worried and forcing me to eat something, however, if I had kept going I would have to assume this would be totally wrong unless I had gorged on warm tea and ACV). Your body will have adapted to creating energy from within and your mind will have come out of the habit of eating every few hours. On day 3 you might even start enjoying it, but not all aspects. Taking naps is a good way to help time past “faster”, and the rest is good for you, too. Being in a low energy state with a severe caloric deficit means you won’t want to waste power on anything so try not to exercise too much (you shouldn’t really at all, but if you must, try to make it something light like a walk or small weights and stretches) Try not to stress out or be in a rush!

Breaking Your 3 Day Fast

It doesn’t really matter when you do it. 70 hours? No problem, your body won’t change much if you cut off 2 hours so no worries. 75 hours? All the same my dude. Want to keep going? Try for 4 days, even 5. Whatever feels suiting, but of course don’t feel obligated to keep going even if you aren’t hungry! There are no nutritional benefits in doing so unless you’re severely overweight or have a medical condition (in which case, please consult a doctor!!!) The absolute worst thing you could do to break a fast is to binge, and worse to binge on junk food! Doing so would stress out your body and namely your intestines. Your body needs time to adjust to food and eating again! Start off with hot lemon water to produce digestive enzymes. ACV will also wake up your intestines and prepare them for food. Before your first actual meal, eat something small and low-glycemic (ex: half an avocado, veggies, nuts). Your first meal should also be small and low-glycemic. This keeps you in a semi-fasted state because of your blood sugar. Carb re-feeding causes abrupt weight GAIN so avoid them like the plague. After your first meal, wait 1-2 hours and continue with small, low-glycemic snacks and meals.

Not eating for three days created a caloric deficit of 6000 calories! However!! This does NOT mean you have lost 6000 calores! Your body is preservative. Don’t eat those 6000 calories back though, obviously. Eat BELOW the normal daily caloric intake! If you eat too much right away, your stomach will be upset. Eat 200-400 calories below your maintaining caloric intake the first day. On the second and third, try ot eat 200-400 above that same number, then adjust your intake to suit your goals! That’s it! You’re all done! I don’t know how to wrap things up other than this soooooo a reminder: please stay safe. I know having an ed or just fasting in general can be really hard so if you ever feel dizzy, no matter how much you want to keep going, please eat something. I had never fasted for more than 24 hours before, so fasting for 48 made me dizzy and feel like I was going to pass out. The funny thing about it is I used to eat all the time in high school, so fasting for 48 hours is unreal to me, but I did it! If you can’t fast for 72 hours right now, it’s no big deal love! Just work your way up to it. If you can only do 12 hours, that’s totally fine! Try for 14 or 16 next time! Just see how much you can do and try to push just a liiiiiitle bit past that point okay? But above all please be safe and take care of yourself!

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Autophagy / オートファジー (“self-devouring”)
Hiiragi Kirai (music, lyrics) / WOOMA (illust, video) / flower (singer)
miy_yuu (cover)
Japanese and English translation included below

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