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Trigger effects on Quirkless - potential risks and possible mutations in cells

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Trigger effects on Quirkless - potential risks and possible mutations in cells



Linda Rose Frankovitch, Mauri Frankovitch, Huyori Akai, Chishiki Sagasu, Madhur Evelyn , Vanna Lakshmi , Isi Michi , Golshan Eun-Jung , Justy Rashmi , Lauren Elian

Department of Quirk Science of Oxford and Tokyo Quirk Research Institute



Summary: Assessing the potential risk Trigger may have on Quirkless - changes in Quirk Factor and binding sites and subsequent mutation, and possible countermeasures.

Methods: Enzyme immunoassay of Quirk Factor concentration. Checking on 3 samples taken from each patient how a certain concentration of Trigger results in changing the cell culture to access potential mutations.

Results: Side effects depends on the type of Quirk written into DNA. There is a high inter-individual variability which unable to generalize. There is hard to predict the exact reaction Trigger may cause in a certain individual due to their Quirkless status and the fact that their body wasn’t accustomed to carrying any Quirk. Trigger for Quirkless posses the same, if not bigger, risk as for people with Quirks.

 

keywords: Trigger, Trigger effects on Quirkless, Quirk Factor, mutations, Quirkless




1 | INTRODUCTION

Trigger is an illegal Quirk enhancing drug which has shown to have a mutative effect on those who take it in high doses or use over a prolonged period. Lately, this drug started to appear more frequently and people dosed with it are constant patients in hospitals. As the problem arose, it became clear that this Quirk enchanting drug may also pose athe risk for Quirkless people who recently started to be a target for Quirk terrorists.

Trigger is a combination of two main components. One is a drug similar to Amphetamines. A central nervous system stimulant that affects chemicals in the brain and nerves that contribute to hyperactivity and impulse control. The second part is a synthetic Quirk Factor. When bound to a normal binding site, the site undergoes a mutation that not only causes the site to shift into a hypersensitive state, it also becomes impossible to shut down the site. As a result, there is oftentimes a permanent mutation of an individual’s Quirk. The other components are unknown, they change rapidly, so stating the exact composition is impossible.

In this study, the pure Trigger will be used.

The potential risk for Quirkless is similar to that for the Quirked population, although the side effects and immunological reaction in Quirkless are unknown, and is based only on speculations. The biggest risk poses the fact that during research conducted on a group of rats, that were prepared to have binding sites and Quirk information coded in their DNA, researchers found out that it is possible for Quirk to appear only once for a short amount of time if the dose of Trigger was high enough.

This review study will focus on analyzing Quirk factor concentration in Quirkless, and the potential mutation Trigger may cause in cells to determine risk and prepare countermeasures.




2 | MATERIALS AND METHODS

2.1 | Study design and patients 

The study was designed in such a way as to collect blood samples from Quirkless individuals from around the world to bridge differences associated with a research group consisting only of representatives of one race, which would significantly narrow down the analyzed gene pool. Data on the genetic background were collected from patients, which included genetic testing of whether they possess genes encoding Quirks and their potential type of Quirk. Information on the type of Quirk possessed by their parents was scrupulously recorded for later comparative analysis.

532 people were qualified for the research group. All of them showed no Quirk, all had an additional joint in their foot. All of the examined group had a check-up under the watchfulness of a specialist whose Quirk made it possible to determine with one hundred percent accuracy whether someone had the Quirk or not. Everyone in the research group received a negative result in that testing. Everyone was Quirkless and their status was confirmed. Candidates who had the Quirks but were lost during their lifetime because of the use of pharmaceuticals, past medical conditions or other causes were excluded from the study. Patients who lacked a foot joint were also excluded from the study due to not fulfilling the stated criteria. All patients in the study group did not have any chronic diseases interrupting the study and were in good health at the time of collecting samples.

The control group consisted of 468 people. The control group consisted of Quirked people, matched to the study group in terms of the types of Quirks possessed by their parents. They were mainly healthy employees of medical facilities and their families who voluntarily agreed to participate in the study. No one in the control group was related to the study group. Individuals applying their Quirk to work were excluded from the study because of the possible risk of fake positive Quirk Factor results due to elevated Quirk Factor level resulting from their professional Quirk usage.

The medical personnel collecting the material and carrying out the entire analysis did not know whether they were dealing with patients with Quirk or not. Only the group analyzing the overall results received this key information.


2.2 | Blood collection, transport of the samples and the way of adding the Trigger

Three 10 ml samples of venous blood were drawn from the ulnar vein (every sample was collected from one vein ) from each patient. The vacuum method of collecting blood was used. Blood was collected in special tubes containing nourishment so that the blood cells did not die during the process of Quirk Factor analysis, and could be cultured to distinguish changes in DNA. The taken samples were cooled to 5°C (41°F) and placed in a cool box whose temperature was constantly monitored. All samples were transported to the Department of Quirk Science of Oxford and Tokyo Quirk Research Institute by teleportation Quirk. One person was responsible for this process to minimize the mistakes made during transporting the samples during the pre-analytical phase. Samples from patients in the vicinity of the Department of Quirk Science of Oxford and Tokyo Quirk Research Institute were also transported using teleportation Quirk to compensate for pre-analytical error associated with uneven sample transport. The teleportation of the samples did not affect the level of Quirk Factor, which was checked in the pilot study preceding the actual analysis. Then all 3,000 samples were refrigerated at 2°C (35.6°F) for twelve hours.

1 ml of blood was collected from all three samples from a given patient to separate plastic probes. After heating samples to 36.6°C (97,88°F), the level of Quirk Factor was tested with the use of enzyme immunoassay kits supplied by two manufacturers - Quirk FactXy6 ENZKT, and Q.Fact001 Chipey. Then 20mg of pure Trigger within a 5ml saline solution was added to the tubes marked with numbers 2 and 3. Samples were left to incubate for two hours for the reaction to start. The Quirk Factor level was measured in all three samples for each patient. Cobis analytical platforms were used to aid in this measurement. Nutritious medium recommended by the manufacturer was added to all tubes to prevent white blood cells from dying. We focused on white blood cells because they have the nucleus which contains the genetic material essential for further genetic testing. The tubes were put in an incubator, where they spent 24 hours at 36.6°C (97,88°F). The next day, the Quirk Factor level was tested before adding a new portion of Trigger. 20mg of pure Trigger within a 5ml saline solution was added to the tubes marked number 3, all tubes were left to stand for 2 hours to incubate with Trigger and then the concentration of Quirk Factor within all the samples was measured. The nutritious medium was added to all tubes. The probes were then stored in an incubator, under the same conditions as before. After an additional 24 hours, the concentration of the Quirk Factor was measured in all samples.

 

2.3 | Tissue culture and gene expression testing

The appropriate amount of material necessary for culture white blood cells on Cul2R growth media was taken from each tube. The blood cell culture was carried out at 37°C (98.6°F) in falcons for 3 days. After three days, the growth rate of the culture was evaluated under a confocal microscope, the medium was changed and further culture was carried out for another 3 days at 36°C (98.6°F). After this time, the growth and appearance of the cells were evaluated visually under a microscope. Colchicine was added to the last 24 hours of incubation to keep cells in metaphase so that the chromosomes are well visible.

A thorough analysis of genetic material derived from white blood cells was performed using a new generation typing. The analysis was performed by experienced genetics with many years of experience, especially in finding changes in the structure of chromosomes, well accustomed to chromosomes originated from white blood cells. The final results reports were based on the latest gene database Ensembl 2218. The expression of genes that were mutated as a result of adding Trigger and the potential impact of the mutations on the life of Quirkless patients was carefully checked.

 

2.4 | Data normalization and statistical analysis 

Data analysis was carried out in the Stati+C program by a qualified statistician. The normality of distribution was tested using the D’ Agostino Pearson test. Differences between patient groups were assessed using the Mann-Whitney U test and Kruskall-Wallis test. To investigate the correlation between two nonnormal continuous variables, a rank test was carried, whereby Spearman’s rho was calculated. A p-value <0.05 was considered a priori to be statistically significant.

Quirk Factor concentration was compared with reference values for the Quirked group, which are in line with international standards for civilians not using their Quirk for their work. Reference values developed based on long-term analysis of patients with Debilitating Quirk Disorder (DQD) were also taken into account to compare with Quirk Factor levels acquired after adding Trigger.

 

2.5 | Ethics statement

The study was approved by the International Quirk Research Union, Department of Quirk Science of Oxford and Tokyo Quirk Research Institute, and hospitals in which the samples were collected. For the safety of patients, the names of certain medical faculties where the samples were taken are disclosed

Written informed consent was obtained from all subjects before sample collection.




3 | Results

 

3.1 | Quirk factor level in subsequently measured samples

In control samples not containing Trigger in the research group, the Quirk Factor level is low and amounts to QF= 1.43 ± 0.5 RQFU (Reference Quirk Factor Units). In control samples derived from a Trigger-free control group, the Quirk Factor level is QF= 235 ± 13 RQFU.

After adding the first portion of Trigger, Quirkless's Quirk Factor level increased to QF =151 ± 32 RQFU, and for Quirk Holders up to QF= 302 ± 18 RQFU. After a night in the thermostat in both groups, the level of Quirk Factor fell by about 23% compared to the intended value two hours after the Trigger administration.

The second portion of Trigger has raised the Quirk Factor in Quirkless to level QF= 248 ± 24 RQFU, and in Quirked control population to the level of QF= 346 ± 7 RQFU. After the night in the incubator, the level in the samples was as follows:

  • In tubes marked with number two, the level dropped by an additional 17% from the initial value after adding Trigger in the control group and by 11% in the test group.
  • In test tubes marked with number three, the level in the control group dropped by 19%, while in the test group by 13%.

It follows that the Quirkless population is more susceptible to maintaining a high level of artificial Quirk Factor in the body compared to the control group in which the body tends to return to the original level of Quirk Factor. Quirkless situation is different. A sudden increase in the level of Quirk Factor is a shock to the body, the second main component of Trigger causes the body to be stimulated and put it into a state of increased metabolism, which is forced and is not a physiological phenomenon. This condition is compounded by additional stimuli resulting from the appearance in the body of a very high concentration of Quirk Factor. The constant high level of adrenaline prevents the smooth decline of new, artificial Quirk Factors. the Quirkless person's body is not used to their high level and therefore there are no functioning mechanisms to remove them from the body. 

 

3.2 | Change in gene expression in cells

3.2.1 | General changes

As a result of Trigger, the expression of genes in stem cells changes, which leads to systemic dissemination of the tumor process. New mutated versions of healthy cells are beginning to spread rapidly throughout the body. The process of their division is accelerated compared to healthy cells. Trigger-added samples contained 103 more cells than the Quirkless controls. For the Quirked, the difference was not so dramatic because of the existing mutations under the control of the Quirk possessor. Cancer changes in the context of Quirk are normal, but not for Quirkless. In the case of Quirkless, the body's efficiency decreases significantly, the cells begin to show the Warburg effect (oxygen glycolysis) - the process of glucose metabolizing to lactate by cancer cells. The body's metabolic capacity decreases, cells become hypoxic. Changes observed in gene expression can translate into systemic changes, which are mainly characterized by increased metabolism, decreased coordination, hypertension, heart arrhythmia, and various heart problems, migraine, permanent muscle twitches/ticks.

As a result of mutations in nucleated cells caused by Trigger, the expression of genes for hypoxia increases significantly, especially the one coding HIF- hypoxia-inducible factor. Disturbing is the change in the expression of genes responsible for the proper functioning of myelin and myelin cells. The study was unable to clearly state the effects that would be expected, but the most likely hypothesis is that Trigger use leads to demyelination of neurons, thereby reducing the rate of transmission of the nerve signal, which translates into a disruption in the functioning of the Quirk, if present. This phenomenon would also explain the degree of dementia people find themselves shortly after taking this drug. They lose their self-control, which is a direct result of the disorder of nerve impulse transport. Trigger has a neurotoxic effect and disrupts the proper functioning of the Peripheral and Central Nervous System.

The spectrum of changes that have occurred in DNA and mutations in chromosomes is very extensive. this is due to various genetic predispositions and latent mutations inherited from the parents of patients from the research group. However, there is a tendency to show similar changes in gene expression in patients with similar genetic backgrounds.

 

3.2.2 | Mutation type Quirk users and Quirkless with the Mutation type Quirk genes predisposition 

Cells of people with parents predominantly endowed with the Mutation (heteromorphic) Type of Quirk more often show changes in the expression of genes encoding epidermal or endothelial cells. In 15% of cases, the genes encoding melanin - the dye responsible for skin color - were mutated. In sporadic cases, there were also mutations in genes encoding the correct division of connective tissue cells and mutations in genes determining the composition of sebaceous gland secretions.

In the case of the control group, increased expression of genes responsible for the manifestation of the phenotype of their mutation could be seen. The changes that could occur with them after direct use of Trigger were developed based on a model taking into account their current ability and the most different gene expression in cells compared to their gene expression profile with white blood cells not affected by Trigger.

 

3.2.3 | Transformation type Quirk users and Quirkless with the Transformation type of Quirk genes predisposition 

A research group displaying a genetic predisposition to the Transformation type of Quirk may experience the greatest complications after trigger administration. It is well known that this group of Quirk holders under the influence of Trigger suffer the most and the mortality rate is the highest. The results acquired during this study showed similar possibilities for Quirkless individuals. Cardiovascular symptoms associated with hypoxia are most severe in them, due to the increased expression of the genes associated with it. Metabolic stress is unbelievably high and cells cannot withstand and undergo necrosis - unplanned cell death. The body may not keep up with its efficient removal from the body, which can lead to anaphylactic shock.

 

3.2.4 | Emitter type Quirk users and Quirkless with the Emitter type Quirk genes predisposition 

Emitter type Quirk users are the most difficult when it comes to analyzing the effect Trigger has on them. They are less likely to suffer from long-lasting side effects. However, they are most likely to suffer the backlash of their Quirk outpacing their bodies. In many cases, they also leave the most amount of damage when they lose control. The variety of changes is the widest and the change in gene expression from blood cells treated with Trigger is the most different from that of healthy cells.

Compared to other types of Quirk owners, the changes were also the most significant in the Quirkless group. The most common mutations regarding the Quirk Factor are mutations in the following genes: QUI1A1, QUI1A2, QUI5D1, QUI5D2, QUI8A1, QUI8C3, QUI8C5, QUI9B5, and QUI9B6, which are the mutations characteristic for Debilitating Quirk Disorder. It may lead to the conclusion that Trigger in Quirkless causes something similar to DQD. Some of the predicted symptoms make this thesis believable. Translocation of 7q21.37 to 17q21.33, where the pathological fusion of these genes occurs, was spotted instantly in 35% of cells. Which is also one of the major etiology factors for DQD.

These symptoms and unusual changes within the genes encoding the Quirk factor binding site were the basis for the hypothesis that says that as a result of an overdose of a trigger, a person with a genetic predisposition to Emitter type of Quirk may manifest symptoms of gaining a Quirk for a short time.



4 | Discussion

Studies in rats have shown that Trigger as a drug causes the destruction of epithelial and blood-brain barrier cells, which leads to leakage and brain damage. The demyelination process is also present. studies using different dosages and trigger concentrations have shown that it has a neurotoxic effect and interferes with the proper transmission of nerve impulses, which can translate into malfunctioning of Quirk if the individual possesses one. This phenomenon is not surprising to anyone who has seen how a person freshly after the administration triggers a regime. Such a person loses his temper and the animal reflexes and primal desires begin to take over. Craving for power, fulfilling your whims and an excessive desire to destroy everything around you. Due to insufficient brain oxygenation, it often happens that a person exposed to Trigger does not have memories of the direct administration of this drug and the subsequent hour. During this time, there is usually the biggest damage associated with the uncontrolled use of the unit's Quirk.

Rats that showed such symptoms also suffered from multi-organ complications that, in individuals chronically exposed to high doses, led to death due to myocardial hypoxia and myocardial infarction.

It would seem that the Trigger problem should not affect Quirkless people at all. Unfortunately, they have recently become a target for terrorists who want to cause even more confusion and contribute to the negative perception of the heroes, because they were not able to protect civilians, especially the most vulnerable according to the public - Quirkless.

The problem is also worrying since, in hospitals and medical units that are the closest to the place where the villains attacked, there are not always personnel trained to react quickly. Trigger intoxication cases are becoming more frequent, and the correct medical assistance in the first hours after the drug enters the body is crucial in the process of rapid recovery.

As a result of Trigger, the expression of genes in stem cells changes, which leads to systemic dissemination of the tumor process. New mutated versions of healthy cells are beginning to spread rapidly throughout the body. The process of their division is accelerated compared to healthy cells. Trigger leads to stem cell damage. Mutations and changes in gene expression arise in a progressive, cascading manner. There is a disseminated systemic neoplastic process. As a result, the individual may for a short time manifest the newly formed as a result of mutation Quirk for a short period. Genetic predispositions to inherit Emitter-type skills are most likely to occur if both parents had Quirks.

Administration of Trigger to a Quirkless person may lead to anaphylactic shock associated with the entry into the body of a foreign protein that is not recognized by the host's immune system. The artificial Quirk factor is not always compatible with binding sites and can lead to an extensive inflammatory process. A very common symptom after administration or taking a trigger is an increase in body temperature.

Low temperatures have proven to be an effective method of combating Trigger. Lowering body temperature by 2 degrees inhibits the spread of the trigger across the body, which significantly facilitates its removal from the body because unbound Quirk Factor is much easier to wash out. Currently, very often used in the therapy of endowed and Quirkless people is cryotherapy.

It follows that the Quirkless population is more susceptible to maintaining a high level of artificial Quirk Factor in the body compared to the control group in which the body tends to return to the original level of Quirk Factor. Quirkless situation is different. A sudden increase in the level of Quirk Factor is a shock to the body, the second main component of Trigger causes the body to be stimulated and put it into a state of increased metabolism, which is forced and is not a physiological phenomenon. This condition is compounded by additional stimuli resulting from the appearance in the body of a very high concentration of Quirk Factor. The constant high level of adrenaline prevents the smooth decline of new, artificial Quirk Factors. The Quirkless person's body is not used to their high level and therefore there are no functioning mechanisms to remove them from the body.

Trigger has a neurotoxic effect and disrupts the proper functioning of the Peripheral and Central Nervous System. As is well known, Quirks are dependent on our nervous system and limbic system, because our Quirks are significantly affected by emotions. From the countless number of collected case studies, one can safely conclude that strong emotions stimulate Quirk owned by a given individual. When strong emotions are manifested, Quirk can activate automatically. It is a kind of defense mechanism associated with the rapid release of a large dose of adrenaline, which puts the body in a state of metabolic stress, to deal with the threat as soon as possible. This is called fight or flight mechanism. Another argument confirming this thesis is also the fact that children begin to manifest their skills at the moment when strong emotions tear them. most often when they are very happy, very bad or feel threatened. There is an incredible number of cases in the literature when the skill came out later than at the age of four. This usually happened in a life-threatening situation or an extremely strong emotional load. (One case described that a woman discovered her skill during her wedding and flooded the church with rose petals). Linking our Quirk control to the limbic system is also justified if one considers the quite controversial but incredibly intriguing theory of Quirks Evolution. It says that under the influence of strong emotions and beliefs, Quirk can evolve automatically to adapt its capabilities to the needs of the user. Many of the heroes who were reporting after a very hard and exhausting mission mentioned that they were able to bring it to an end only because of the sudden surge of power from their Quirk. The heroes pointed out that they had never encountered this before and that after this episode their Quirk seemed stronger.

Trigger is a drug that can lead to short-term memory loss, usually, it is the time after taking the drug and this phenomenon lasts from 10 to 30 minutes, depending on the capacity of the nervous and immune systems of the user and the dose and purity of the drug introduced into the body. Prolonged use of Trigger can contribute to permanent damage to the psyche and personality disorders, as well as dysregulation of the punishment and reward system

It is difficult to predict how an individual will respond to this drug. In our population, there is a very high variability associated with a huge number of spontaneous mutations, thanks to which we can use the Quirks owned by the majority of the society. Even Quirkless, having no special skills, have a genetic predisposition associated with a particular type of skill. Symptoms after administration to the system trigger are strongly associated with these genetic predispositions.

Depending on the dose used, Quirkless with Mutation type Quirk genes predisposition may experience a progressive mutation that may change their external appearance or reorganize internal organs. This condition is extremely dangerous because the proper physiology of these pathological changes will be maintained for a very short time and the person who experiences it will suffer from huge pain and in the worst-case surgery will be necessary. to restore the body of a person exposed to Trigger to its original state.

Quirkless with the Transformation type of Quirk genes predisposition is the group most at risk of complications. Even a small dose of Trigger introduced into their system can lead to anaphylactic shock and a radical defensive reaction of the body. In the worst case of long-term use of large and contaminated doses of this organism, death will occur as a result of cardiac arrest due to hypoxia of the blood supplying vessels, or cerebral death as a result of cerebral hypoxia due to cerebral arterial clots.

Quirkless with Emitter type Quirk genes predisposition on a single exposure to small doses of Trigger, the risk is not high. It is worrying that in this group the most mutations occur in the genetic material. there is a risk of developing DQD, which would seem absurd in a Quirkless person. Unfortunately, genetic research supports this hypothesis. As a result of administering a very high dose, there is a risk that a person will be able to represent a Quirk for a very short period before the rapid depletion of the body. This can lead to multi-organ damage and even death.



The authors are indebted to the blood collecting nurses assisting in the process in hospitals participating in this big project.

The authors have declared no conflict of interest.

The further study was prohibited by International Quirk Research Union due to the potential risk the research may be caused to the patients.