The ketogenic diet, commonly referred to as ‘keto,’ is an increasingly popular recent trend in the dieting world. While there are a variety of ketogenic diets, including the Atkins and South Beach diet, the principle idea behind keto is that you use fats and ketones in your body to provide energy, rather than carbohydrates and glucose.
However, keto isn’t just a new practice that started in the past couple of years. With the first records of this practice dating back to the time of Hippocrates, ketogenic diets and fasting have been used for many medicinal reasons besides weight loss such as helping to treat or reduce the symptoms of various conditions such as acne, polycystic ovary syndrome to even genetic disorders.
Ketogenic diets have been studied extensively in people with epilepsy. They are often recommended for those who either fail to respond to anti-seizure medications or can’t tolerate their side effects and have been proven quite effective.
How Does the Ketogenic Diet Work?
Typically in standard diets, some carbohydrates burn up quickly and are packed with intense fuel that yields large bursts of energy. The human body’s main energy store is through fats . Proteins and fats burn slowly thus results in a constant stable release of energy. This is beneficial as you would not experience any peaks or crashes in energy in comparison to carbohydrates by which glucose is released instantaneously in the body and not stored [1,2]. This is how ketosis also regulates blood glucose levels without complications. Involving complex carbs into your diet causes your body to heighten your blood sugar and as a result, produce insulin. As high glucose levels are perceived as toxic by the body, insulin is released by the pancreas as negative feedback to decrease and regulate this. When your body starts struggling to keep up, excess glucose is converted into fat, and insulin stores it in cells. Your body is capable of regulating your blood sugar on its own without help when you aren’t mainlining so much sucrose. Ketogenic diets avoid such problems.
Fat is essentially satiety. Consuming fat keeps you full and combines with low carbohydrate intake, allows the body to produce ketones [2,3]. In Glucose 1 transporter (Glut 1) deficiency and pyruvate dehydrogenase (PDH) deficiency ketones provide an alternative energy source to glucose .
As with any alterations to the body, the Ketogenic diet too has side effects whilst adjusting to the low-carb lifestyle. Though for many the intended goal is weight loss, in other cases where the ketogenic diet is prescribed, side effects can include feeling weary with symptoms of influenza, also known as the Keto-Flu, constipation, or diarrhea, and high cholesterol.
The Ketogenic Diet for Epilepsy
Most often lifelong, epilepsy is a common condition that affects the brain and causes frequent seizures. Seizures that are resistant to standard medications remain a major clinical problem.
In severe epileptic patients, after 2-3 days of fasting from food, and even in some cases, water, there was severe seizure control observed. These observations have been around for millennia.
We don’t completely understand how the ketogenic diet works but there is some evidence that the brain needs energy from glucose to create a seizure. The ketogenic diet makes the body think that it’s in a state of starvation or fast .
Though having years of insight and trials, clinically, doctors do not assign patients keto from initial diagnosis of epilepsy; it is usually reserved for drug-resistant epilepsy. This is due to the balance of the diet needing to be carefully worked out for each individual and because vitamins and other special supplements are needed. It is not advisable as a treatment option for some with some metabolic disorders or other neurological disorders.
The ketogenic diet was initially for children and currently in the UK, medically still is; this treatment is offered for children between 3 months to 16 years suffering from any type of epilepsy [2,4,5]. During infancy, the brain is much more efficient at extracting and utilizing ketone bodies from the blood due to the higher levels of ketone metabolizing enzymes and monocarboxylic acid transporters produced during this period [6-8].
In a review by Mcgill et al., four studies with 385 participants reported that more children achieved seizure freedom with a ketogenic diet than with control. Two small underpowered studies reported no events in adults with 141 participants reported no adults achieved seizure freedom; the analysis was underpowered .
There is limited evidence for how effective or tolerable the ketogenic diet is for adults but it has been highly proven to be effective for pediatric epilepsy. Some particular studies show that keto can be effective on the adult brain as it increases its levels of monocarboxylic acid transporters and ketone metabolizing enzymes rapidly during periods of stress such as ischemia, trauma, or low glucose as in the case of keto [8,10]. A review of 16 studies in adults with uncontrolled epilepsy found that ketogenic diets were well-tolerated long term and typically resulted in significantly fewer seizures or, in a minority of cases, complete freedom from seizures .
A study by Rho et al. suggests that adults may produce ketones at different levels in response to a ketogenic diet, possibly explaining the inconsistencies in findings on the effect of keto on the adult brain. In addition, they also demonstrated variations between the production of ketones in rats and humans, which is a crucial consideration when running animal trials before implementing techniques and diets clinically .
The ketogenic diet is not just a trend for health gurus to lose weight. It lessens the burden and potentially frees many, if not, millions of those suffering from drug-resistant seizures from epilepsy. Though not much is known about epilepsy as a condition in itself, and the ketogenic diet heavily varying in effect amongst people of different ages, for centuries, techniques like fasting and keto have been observed to be helpful. Though not as commonly prescribed or allocated to adults, it is an extremely effective technique on children, potentially improving their trajectory and progress of the condition.
Nara Ito, Youth Medical Journal 2020
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 Lambrechts, D. A., de Kinderen, R. J., Vles, J. S., de Louw, A. J., Aldenkamp, A. P., & Majoie, H. J. (2017). A randomized controlled trial of the ketogenic diet in refractory childhood epilepsy. Acta neurologica Scandinavica, 135(2), 231–239. https://doi.org/10.1111/ane.12592
 Sharma, S., Goel, S., Jain, P., Agarwala, A., & Aneja, S. (2016). Evaluation of a simplified modified Atkins diet for use by parents with low levels of literacy in children with refractory epilepsy: A randomized controlled trial. Epilepsy research, 127, 152–159. https://doi.org/10.1016/j.eplepsyres.2016.09.002
 Bilger, A., & Nehlig, A. (1992). Quantitative histochemical changes in enzymes involved in energy metabolism in the rat brain during postnatal development. II. Glucose-6-phosphate dehydrogenase and beta-hydroxybutyrate dehydrogenase. International journal of developmental neuroscience : the official journal of the International Society for Developmental Neuroscience, 10(2), 143–152. https://doi.org/10.1016/0736-5748(92)90042-x
 Martin‐McGill, K. J., Jackson, C. F., Bresnahan, R., Levy, R. G., & Cooper, P. N. (2018). Ketogenic diets for drug‐resistant epilepsy. Cochrane Database of Systematic Reviews, (11).
Prins M. L. (2008). Cerebral metabolic adaptation and ketone metabolism after brain injury. Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism, 28(1), 1–16. https://doi.org/10.1038/sj.jcbfm.9600543Prins M. L. (2008). Cerebral metabolic adaptation and ketone metabolism after brain injury. Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism, 28(1), 1–16. https://doi.org/10.1038/sj.jcbfm.9600543
Since its discovery in 1973, huge strides have been made in genetic engineering. Some examples include arctic apples, mouse ear cress, and even onions that do not make you cry. Genetically modified food even comprises much of what we eat today, with roughly 60-70% of processed food in grocery stores containing genetically modified parts. However, some are still concerned about the potential consequences of this relatively new technology, as there are unknown side effects. Even though we constantly hear disturbing reports about genetically modified food, we barely hear something substantive about this exciting issue. Researchers have been looking into altering the genetic information of contagious diseases. These diseases have plagued the earth for decades, taking millions of lives along the way. Could the power of genetic engineering be used to eliminate them completely?
The Zika virus has spread exponentially across the globe, through mosquitoes. The virus causes horrific birth defects in pregnant women and weakens people’s immune systems by damaging nerve cells. There is no treatment available. Another virus that spreads in a similar way is malaria. In the salivary glands of a mosquito, thousands of sporozoites wait before the mosquito penetrates a humans flesh, they then move to the liver, while avoiding the immune system. They remain here in stealth mode for up to a month, eating the cells alive and creating thousands of copies of themselves. These diseases are a threat to nearly half the world’s population, and account for millions of deaths. What if you could use genetic engineering to stop the spread?
What is Genetic Engineering?
Genetic engineering, or gene modification, is the DNA alteration process in the nucleus of an organism. According to the national Human Research Institute, genetic engineering can be done by using recombinant DNA (rDNA) or DNA derived from two or more distinct species and then combined into a single molecule.This technology has been used to create things like safer lithium-ion batteries and crops such as Sweet Plum. Genetically engineered plants, known as genetically modified organisms (GMOs), may be engineered to be less susceptible to disease or to suit particular environmental requirements. In addition to this, there are many genetically modified organisms, for example, mosquitoes that were modified in the lab.
How Can This Help Stop Diseases?
The particular virus of Zika is spread through the bite of a mosquito. The mosquito is the natural carrier for human pathogens that have been present for nearly 200 million years. There are trillions of them, and one will lay up to 300 eggs at a time. They’re virtually difficult to eliminate. In order to stop the mosquito the entire mosquito population must be re-engineered. This can be done using a new technology, CRISPR. CRISPR is an easy but efficient method for genome editing. It helps researchers to quickly alter DNA sequences and change the role of genes. It also serves as a tool to modify genes. This can be achieved by adding a cut or break in the DNA and by tricking the cell’s own DNA repair mechanisms into making the improvements that one needs.Through this genetic modification, scientists have successfully developed a strain of mosquitoes that are resistant to the malaria parasite by inserting a new antibody gene directly targeting plasmodium. This same theory could be applied to the Zika virus, or any other similar virus that relies on a host such as Lyme disease, sleeping sickness, and West Nile virus. These mosquitoes would never even carry the disease and millions of lives could be saved. On top of this, if the new gene becomes dominant over the next generation it will overpower the old gene. If sufficiently modified mosquitoes were to mate with natural mosquitoes, the gene would spread very quickly. Thanks to this, 99.5 percent of all designed mosquitoes’ offspring will not carry the virus.
There are still many concerns considering that CRISPR is a new technology. Critics of genetic engineering are vocal about the potential consequences such a move may have, such as potential environmental harm. Additionally, after the gene is edited, there is simply no way to change it back to normal; meaning that those consequences are there to stay if something goes wrong. There are simply many things that we do not know yet. Perhaps the worst-case situation here is that it does not work, or that the parasite adapts in a negative way. Finally there are ethical concerns, with people stating that genetic engineering is “playing with nature.” All of these factors make it unsure if this technology will be used.
Ellen Weiss noticed uneven, desiccated patches on her toddler’s dark brown skin and wondered if the patches signified eczema, or rather a skin condition more serious. “I Googled it and noticed immediately the pictures were all of white skin. I Googled other conditions and it was the same. No matter what I searched, there were almost no images of dark skin,” she recounted (McFarling).
The availability of medical reference photos, particularly those depicting dermatological conditions on dark skin, is an issue that has not received the awareness it needs. Today, many Brown and Black individuals encounter problems identifying a condition on their skin. Even when approaching a doctor, people still do not receive the help that they need (Prescod). The inherent problem concerning medical images is the inequality of reference material from across the skin tone spectrum, necessary for care and treatment of their skin’s health.
Medical images are crucial in training future physicians as these images display the anatomical and etiological aspects of the body. With such material for reference, physicians are able to recognize certain diseases, disorders, and injuries a patient may have. For instance, an assessment of the intensity of burns is dependent upon a physician’s visual inspection (Schaefer). By looking at certain characteristics of a burn, doctors can identify the injury as one of three measures: first degree, second degree, and third degree. Burns are only an example of which its primary method of diagnosis is through visual inspection alone, and as this process enters the realm of dermatological conditions, it encounters complications that obstruct the way for an easy, and sometimes correct, diagnosis for individuals with dark skin (Singh).
Before investigating the hardships when it comes to diagnosing dermatological conditions on dark skin, it is imperative to first understand the inherent statistics of medical images concerning the measure of racial diversity that they encompass.
Rhiannon Parker, a researcher at the University of Wollongong in Australia became aware of the lack of diversity in medical images. Parker and her colleagues sought to understand the extent to which the lack of diversity in dermatological reference photos exactly accounts to. Inspecting more than 6000 medical images derived from 17 different anatomy books published between the years 2008 and 2013, they firstly focused on gender disproportionality. They found that only 36 percent of images with an identifiable sex were female, and they furthermore analyzed these images’ racial make-up. Of these images that displayed females, 86 percent of them were White (Eveleth).
A more recent study, conducted in April 2020 and led by Jules Lipoff, an assistant professor of clinical dermatology at the University of Pennsylvania, analyzed the skin type diversity in general medicine textbooks. Lipoff found that only 4.5 percent of images with an identifiable skin tone displayed dark skin (McFarling). The disproportionality within medical textbooks is evidently overwhelming.
Dermatology is a branch of medicine that focuses on the structure, etiology, and treatment of the skin. Dermatologists undergo years of training to especially develop an ability to diagnose a certain skin condition based on visual inspection. However, dermatologist Jenna Lester from the University of California, San Francisco describes the difficulties she encountered when preparing to treat her Black patients for a particular rash commonly associated with Covid-19. The popularly known “Covid-19 rash” is an emerging symptom of coronavirus that scientists are still researching. When Lester heard of this rash, she searched throughout various medical literature to analyze what the rash would particularly look like on dark skin. From her ongoing searches, Lester still did not find one picture displaying the rash on dark skin (Lester).
She mentioned, “I was frustrated because we know Covid-19 is disproportionately impacting communities of color. I felt like I was seeing a disparity being built right before my eyes” (McFarling).
Considering the emerging idea that certain dermatological conditions can look very much different across the skin tone spectrum, the lack of racial diversity poses disadvantages for individuals with dark skin (Figure 1). As physicians continue to train upon a white foundation, those with darker skin face errors and complications made by doctors within inspection, and therefore face higher rates of misdiagnosis (Rabin). The hardships that many encounter when simply attempting to identify a skin condition is beyond a representation of today’s inequality for people of color.
This issue is relatively new and merits a call for social awareness. For instance, Malone Mukwende, a second year medical student at St. George’s University of London, strives to combat the inadequate research in dermatological conditions on dark skin.. He and his colleagues are working together to write a book, “Mind the Gap,” which addresses the various clinical signs of skin disorders on black and brown skin (“A Medical Student”). At the time of this article, the book is not yet published, and still no official release date has been confirmed. Furthermore, a book by Cyron Mandia, “All About the Skin: A Microscopic Lens to the Integumentary System,” which will be published around this December, utilizes its incorporation of dark skin to combat the racial misrepresentation within dermatological images.
As society continues its works to fight the lack of diversity in medical images, a modern and racially inclusive dermatological curriculum waits upon us. However, until then, it is our duty to continue to spread awareness for change.
Lester, J. C., Jia, J. L., Zhang, L., Okoye, G. A., & Linos, E. (2020). Absence of images of skin of colour in publications of COVID‐19 skin manifestations. British Journal of Dermatology, 183(3), 593–595. https://doi.org/10.1111/bjd.19258
Artificial organs are a recent form of life saving technology that rely on methods such as 3-D printing and stem cell implementation, which are becoming more common in the medical community as automation advances. Lately, scientists have been putting in a lot of research and resources into the development of these artificial organs, particularly as a result of the significant imbalance between those in need of organs and those who are actually receiving them. In 2015, about 121,000 people needed an organ transplant, but only about 31,000 patients received one (Figure 1).
Patients that Received Organs (blue) vs. Patients in Need (grey)
Figure 1, Organ Procurement and Transplantation Network, 2015
This gap is a huge problem; people with failing organs do not have much time to spare. According to the Nation Health Resources and Services Organization, about 20 people die on average per day because of a lack of viable organs for transplant. As a result, artificial organs have become especially important, as they provide other means for people to obtain the organs that they require.
A Societal Perspective
The wait for essential organs is absurd and inhumane. In fact, it can generally take up to five years to get off the waitlist for a transplant, which can cost a human life. Consequently, some find it necessary to resort to unconventional ways to get what they need. As expressed by Nancy Scheper-Hughes, a professor of anthropology at the University of California, Berkeley, the extensive wait for these organs often motivates people to take the state of their lives into their own hands. In this instance, they become willing to travel long and far to obtain organs that they receive through channels of legal or illegal means. These distances are often foreign countries, with the most popular destination being India.
With the implementation of biomedically engineered artificial organs, these desperate patients will no longer need to put themselves and even their families in jeopardy to live. This way, the patients are able to receive the organs that will save their lives in a legal way without the trouble of traveling in their unfortunate conditions. With new and improved organs, not only will people have a better quality of life, but they’ll be able to contribute to society in ways that may not have been possible for them before.
The Ethical Dilemma
While these organs may seem like a perfect solution, there are some serious ethical consequences that arise as a result of increased implementation. A main issue that will accompany the spread of these high-tech organs is their large cost. As shown in Figure 2, the cost of procedures that implement artificial body parts such as hips and knees are already what many would consider high, with a simple ear tube procedure costing up to $4,500.
The Cost and Annual Revenue of the Artificial Organ Market
Considering these organs would only be available to those who could pay, it could be considered unvirtuous to allow them to become very widespread, as others that cannot afford these situations will be at a major disadvantage that could potentially decide their quality of life. Also, since one cannot control their health if they are on the verge of death, it becomes immoral to make these organs available to those that can afford the high expenses.
After evaluating this complex issue from multiple perspectives, it becomes clear that a solution is needed. Overall, in order to help solve the issue of organ scarcity whilst reducing the ethical implications, highly-demanded artificial organs should be implemented, however the complexity of their software should be reduced. The limitations of this solution, however, are that it takes extensive research and time in order to maintain the effectiveness of these organs while simultaneously reducing their complexity, and that the artificial organs being produced would only be available for those in need of major organs. Despite these drawbacks, the outlook for biomedically engineered artificial organs is exceptionally bright, and this market has the potential to save a myriad of lives that would otherwise be lost.
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Vermeulen, Niki & Haddow, Gillian & Seymour, Tirion & Faulkner-Jones, Alan & Shu, Wenmiao (Will). (2017). “3D bioprint me: A socioethical view of bioprinting human organs and tissues. Journal of Medical Ethics. 43. medethics-2015. 10.1136/medethics-2015-103347.
Artificial intelligence in medical practice is the use of computer techniques to perform clinical diagnoses and suggest treatments in medical areas . It has the capability of detecting meaningful relationships in a data set and could be used for the diagnosis, treatment, and for coming to a particular conclusion. Similar to the way doctors are educated through years of medical schooling and learning from mistakes, artificial intelligence algorithms learn to do the same job as a doctor. They perform tasks requiring human intelligence like pattern and speech recognition, image analysis, and decision making. The Artificial Intelligence algorithm includes feeding data in the computer system, which are structured having a label recognizable to the algorithm, performance is analyzed just like exams give an analysis of a medical student’s performance thus giving results. Based on the results of this analysis the algorithm can be modified, fed more data, or rolled out for the decision-making of the person writing the algorithm .
Fig: AI algorithm learning the basic anatomy of a hand and can recreate where a missing digit should be. This could allow physicians to see the proper place to reconstruct a limb/put a prosthetic.
These performances and results are tested with a physician’s performance to determine its clinical ability and value. In medicine language, it includes input data based upon numericals such as Heart Rate or Blood Pressure and based upon images such as Magnetic Resonance Imaging Scans or Images of Biopsy Tissue Samples. The algorithms from this data could be a probability or a classification. The result of the above example could be the probability of having an arterial clot according to the heart rate and blood pressure data or the labeling of an imaged tissue sample by classifying it as cancerous or non-cancerous. There are two recent applications in the Artificial Intelligence of clinical and accurate algorithms benefiting both patient and doctor for the diagnosis. One is the algorithm researchers at Seoul National University Hospital and College of Medicine developed called Deep Learning-based Automatic Detection to analyze chest radiographs and detect abnormal cell growth (cancers). The results were compared to many physician’s detection abilities and were found to perform better than the doctors .
Fig: Artificial Intelligence Algorithm. Left panel showing the image fed into an algorithm. The right panel shows a region of potentially dangerous cells, as identified by an algorithm, that a physician should look at more closely.
Fig: Artificial Intelligence algorithm; Deep Learning Method.The left panel shows the original X-ray. The right panel shows the X-ray with orange color indicating signs of pneumothorax which could be unnoticed by radiologists
The second algorithm was developed by researchers at Google AI Healthcare called Lymph Node Assistant which analyzed histology slides stained tissue samples to identify metastatic breast cancer tumors from lymph node biopsies. It could identify suspicious regions of the sample which could not be distinguished with the human eye. It was proven to accurately classify a cancer as cancerous or non-cancerous in 99% of the cases. Hence these algorithms could help doctors with correct diagnosis thus allowing them to invest time in solving cases that computers cannot solve .
Fig: AI algorithm; Lymph node biopsy
Artificial intelligence could be considered a boon as it may help for early diagnosis of diseases whose later diagnosis can cause delays in the treatment and may be harmful to the patient. For example, researchers have claimed that it could be used to diagnose Alzheimer’s disease years before symptoms appear. The computers can be trained for brain scans to be able to spot subtle signs of dementia that could be missed by humans allowing early diagnosis. This could probably be done using 18-F-fluorodeoxyglucose positron emission tomography (FDG-PET). In an FDG-PET scan FDG, a radioactive glucose compound is injected into the blood. PET scans can then measure the uptake of FDG in brain cells, an indicator of metabolic activity. Through Deep Learning, the algorithm can teach itself metabolic patterns that correspond to Alzheimer’s disease.If one can detect the symptoms earlier, it would help investigators to find better ways to reduce or halt the disease process. Future research should take into consideration, training the deep learning algorithm to look for patterns associated with the accumulation of beta-amyloid and tau proteins, abnormal protein clumps, and tangles in the brain that are markers specific to Alzheimer’s disease, according to UCSF’s Youngho Seo, Ph.D., which can add another dimension to using Artificial Intelligence in Alzheimer’s disease detection .
Fig: Fluorine 18 fluorodeoxyglucose PET images from Alzheimer’s Disease Neuroimaging Initiative set preprocessed with the grid method for Alzheimer disease patient
Artificial Intelligence has many clinical applications to improve patient care and potentially save lives. Maintaining medical records and past history is the first step in health care where robots collect, store, reformat, and trace data to provide faster and more consistent access. They also analyze data including notes and reports from a patient’s file and clinical expertise to help to choose the right treatment pathway .There are some latest tools and technology developed in the health care sector based on the Artificial Intelligence algorithm. This includes: MelaFind, which is a tool that does not involve introduction of instruments into the body and gives extra information to dermatologists in early detection and recognition of skin cancer,lesions and helps in it’s examination. It also helps in evaluation of skin lesions up to 2.5 mm beneath the skin. By using Artificial Intelligence based algorithms, dermatologists can analyze irregular moles and diagnose serious skin cancers such as melanoma. The device demonstrated 98.3% sensitivity by correctly identifying 172 out of 175 melanomas and high-grade lesions. Robotic-assisted therapy is used in neurological patients and is specially used for stroke patients’ recovery. The robotic arm and hand use digital algorithms to detect motions that patients cannot execute during therapy thus improving their performance per hour than they would have if worked with a physical therapist alone thus allowing speedy recover.Robots can also perform tests, x-rays, CT scans, data entry, and other tasks faster and more accurately. Cardiology and Radiology are two fields where the amount of data to analyze is huge and time-consuming. Future cardiologists and radiologists should look only at the most critical cases in which human monitoring is useful . Caption Guidance, which is an Artificial Intelligence guided ultrasound platform or software capable of instructing clinicians on obtaining a clearer picture of the heart in motion. It will be used for capturing echocardiographic images of the patient’s heart without special training, spotting high-quality 2D heart images, and automatically recording video clips for later analysis, while calculating heart function measures thus improving the diagnosis of heart diseases .
Fig: AI tools in health care
Conclusion: Artificial Intelligence will surely improve the healthcare industry, from predictive medical care and more accurate diagnosis to motivating the patients to take care of their health. It will certainly continue enhancing the patient’s experience and healthcare expertise in general. The use of Artificial Intelligence is predicted to decrease medical costs as there will be more accuracy in diagnosis and better predictions in the treatment plan as well as more prevention of disease. It will not replace healthcare workers but instead allow them to spend more time for the bedside care of their patients, resulting in the greater outcomes for all.
Plastic surgery is often associated with cosmetic procedures, relating to the elective enhancement or change of one’s body or facial features. In the modern age, this has been popularized by social media influencers. Thus, plastic surgery has taken on a reputation of being non-vital and less important than other fields like neurosurgery or cardio-thoracic surgery. However, plastic surgery is not limited to this. Instead, reconstructive surgery is another vital segment.
Reconstructure plastic surgery, on the other hand, is not cosmetic. It is completed on patients suffering from conditions and complications such as cancer, trauma, and deformities. This is meant to normalize their appearance or improve bodily function. Reconstructive plastic surgery is considered to be extremely diverse with many new innovations. Technology, such as augmented reality or the use of three-dimensional imaging is an important and useful part of reconstructive plastic surgery [1,2].
A Japanese research group created an augmented reality system, allowing surgeons to create a three-dimensional version of the results over the original. This involves the use of smart glasses, which project the image over a patient’s face during the procedure. This was designed to be simple and efficient, guiding the surgeons and ideally minimizing mistakes [3,4].
More recently, researchers at The University of Michigan created new technology that can aid surgeons in complex reconstructive microsurgery cases. Known as the “arterial everter” this device with a pen-like structure can essentially make the process of connecting arteries more efficient and simple. The amount of time saved with this device is not only beneficial to the patient, but to doctors and to the healthcare system as well. To provide a scenario, if a patient’s arm is heavily severed due to an accident, this device can be extremely beneficial to the surgeons in this case.
At UC Davis, plastic surgeon Dr. Michael Wong, has tested the benefits of using a high-tech camera in certain surgical cases, such as breast reconstructions. This device generates three-dimensional images allowing patients to see how their end result might look like. The camera is big in size and has multiple arms and lenses which take pictures from different angles, and creates a 3-D rendering through the use of computer software. Not only does this technology allow patients to be more satisfied, but it also allows surgeons to improve the quality of their work.
The youngest patient to undergo a facial transplant–Kattie Stubblefield–had suffered a gunshot wound. The majority of her face was destroyed, and she experienced severe brain injury. After 31 hours, a team of 40 surgeons, and 2 operating rooms, the Cleveland Clinic finished Kattie’s surgery, which is known to be the most substantial face transplant. The procedure required the replacement of all her facial tissue including the scalp, eye sockets, facial muscles, and other components. The primary plastic surgeon working in this case—Dr. Brian Gastman, MD detailed how complex these facial procedures are. The facial tissues from the donor were cut so as to retain function while being transferred to the patient. Then, augmented reality allowed her surgeons to know where to cut and to improve the placement. The use of augmented reality, in this case, comes with many benefits. According to Dr. Gastman, this technique allowed doctors to create ideas in advance and do more extensive modeling.
Overall, technology in reconstructive plastic surgery has proven to be extremely beneficial, as it allows for fewer mistakes, better results, patients are more likely to be satisfied, and it can even make for a better teaching tool.
Scientists attempting to grow human organs themselves are not an entirely new concept; in fact, scientists have been conducting this process for well over a decade, growing human organs ranging from kidneys to skin. Researchers have now been able to grow a mini-brain with neural activity that mirrors a preterm infant. This is an enormous stride forward as earlier work was unable to demonstrate brain activity that was similar to how the brain actually functions.
This research was largely founded based on neural oscillations, rhythmic brain signals found across species. Neural oscillations are one of the many cellular networks that eventually develop into circuits in the human brain during the maturation process. While this eventual development of neural oscillations into circuits is known, it is unclear when these networks precisely develop. Recent students in mice have demonstrated that oscillations develop immediately after birth. However, due to the fact, there previously were not any adequate models of the human brain in the laboratory setting it is unclear whether the same course of events occurs in humans brains.
The team at the University of California, San Diego first grew human induced pluripotent stem cells (iPSCs), cells that can self renew, in neurons found in the cortex of the brain, which is responsible for controlling thought and behavior. The researchers opted to use iPSCs during this experiment as they have been shown in recent studies that they have the capacity to mimic various development features and the cellular and molecular processes of the human brain. In order to successfully grow the pluripotent stem cells, researchers created a solution that contained a mixture of transcription factors that regulate fetal development. Creating the perfect mixture was pivotal as it allowed for organoids to last long amounts of time. Many of these organoids were still usable after a year since the research was completed. These conditions allowed for the researchers at UCSD to mimic the neural oscillations and electrophysiological network activity.
This process differed from earlier attempts to create mini-brains as the team created the most optimal conditions for mini-brain development at every stage. For example, unlike standard protocols which start with a clump of cells to construct the organoid, the team started from a single cell to build the organoid. In addition, they also changed the timings and concentrations of certain aspects in the mixture. While the process was meticulous, it certainly paid off.
The results of this study revealed that the mini-brains produced neural activity that mirrors a preterm infant; thus, mini-brains have the potential to serve as laboratory models for studying psychiatric conditions. The iPSCs used in the study, like most stem cells, can be differentiated to specialize in any cell; in this case, the iPSCs were directed to specialized into neurons and glia.
Due to the team’s unique way of creating an organoid from scratch, the team was able to identify a particular neuron that has previously never been able to be created in a laboratory setting: the GABAergic neuron. Another example of the team’s diligent process paying off was seen when measuring the electrical activity of the mini-brains using an electroencephalogram (EEG). The test revealed the mini-brains following the new protocol had 300,000 electrical impulse spikes per minute. With the old process, the earlier mini-brains produced a mere 3,000 spikes per minute.
The results are what lead the UCSD to compare their results with the electrical patterns in newborn baby brains. The neural oscillations discussed earlier in the introductions, change according to age. Newborn baby brains tend to be the least active in oscillations. Their brains tend to have almost no waves between spikes of electrical activity. As we get older, these periods with no activity tend to get shorter, and eventually become constant. Other than age, oscillation patterns can also be affected by cognition skills and various diseases.
The team compared their mini-brains with a previously published dataset of 567 EEG recordings from 39 babies born prematurely (between 24 and 38 weeks gestation). This cross-analysis revealed that the organoids revealed similar patterns in their EEG levels for up to 9 months after being developed.
The successful development of a functional mini-brain has far-reaching applications as it broadens the range of neurologic conditions that could be adequately studied. Further, in most psychiatric conditions the neuronal circuitry is impaired: mini-brains would allow for a better understanding of diseases such as autism and epilepsy. Prior to this study, there were no adequate models in the laboratory to study certain neurological diseases adequately. For example, Alysson R. Muotri, Ph.D., one of the lead scientists of the study, reported sending the brain organoids to the International Space Station, in an attempt to determine the effect microgravity has on brain development. Potentially, the results yielded from this experiment could determine the prospects for human life outside of Earth.
While developing mini-brains further holds the potential to change the way we study neurological diseases, this study exists in a fine line between science and ethics. Critics of this study and other members of the scientific community ask, “Are we getting too close to re-creating the human brain?”. Muotri understands these concerns and responds to these concerns by stating that the mini-brains developed in the lab are far from being functional adult human brains. He points out that the mini-brains are not only much smaller than fully developed brains but also lack hemispheres and blood vessels. Muotri also stated, “They are far from being functionally equivalent to a full cortex, even in a baby… In fact, we don’t yet have a way to even measure consciousness or sentience…”. Muotri asserts that science is far ways from creating a fully functional, developed brain, but as the medical landscape changes faster than ever ethics dilemmas that arise similar to this situation cannot be ignored.
1. “Lab-Grown “Mini Brains” Can Now Mimic the Neural Activity of a Preterm Infant.”
Introduction: What is CRISPR/Cas9 and How Does it Work?
Clusters of regularly interspaced short palindromic repeats, commonly known as CRISPR, is a simple yet formidable implement allowing researchers to effectively edit genomes. Some of its potential applications include treatment and prevention of diseases, amelioration of genetic defects, and removal of certain gene types. The CRISPR/Cas9 technology can be deconstructed into four components (CRISPR, spacer, crRNA, and Cas9), each serving its own purpose. The CRISPR is a unique sequence of deoxyribonucleic acids (DNA) consisting of two distinct attributes: recurrent nucleotide progressions and spacers, which are short-variable sequences interspersed between nucleotides . These spacers are derived from the DNA of viruses that have previously attacked the host CRISPR . Once the CRISPR has been fashioned and the virus attacks again, the CRISPR is transcribed and processed into crRNA. The crRNA integrates with a secondary RNA string, the trans-activating crRNA, to provide the Cas9 enzyme with a passage to its target site, where the Cas9 enzyme will then execute what is recognized as a double-stranded break to remove the superfluous module of the genome sequence. One theory as to why CRISPR/Cas9 technology is so attractive compared to other genome editing techniques to researchers and consumers alike is its meticulous accuracy and efficiency in deriving results. It is predicted that the rationale behind this lies in the technology’s prototype adjacent motifs or PAMs. These PAMs serve as a tag, sitting adjacent to the Cas9 enzyme’s target site. Should there be no PAM next to the target site, the Cas9 enzyme will avoid “cutting” the particular genome sequence . Once the cut has been finalized, programmed DNA may take its place.
Why CRISPR: A Deeper Understanding
Over the past decade, four major classes of engineered nucleases have been used for genome editing: meganucleases, zinc-finger nucleases (ZFN), transcription activator-like effector nucleases (TALEN), and CRISPR [6.] Meganucleases are endonucleases that can recognize extended DNA progressions of 14-40 bp via extensive non-modular protein-DNA connectors, but their target specifications have proven to be difficult to re-engineer . Both ZFNs and TALENs are “fusions between arrays of ZF or TALE DNA-binding domains and the non-specific, dimerization-dependent FokI nuclease domain” . Put simply, these three classes of nucleases all rely exclusively on protein-DNA connections to recognize their target sites.
In contrast, the CRISPR/Cas9 endonuclease relies on RNA strands to guide the Cas9 protein to its target site. This process allows researchers to easily modify and reprogram gene sequences. Additionally, due to the Cas9 enzyme’s complex nature of being intersected with a secondary RNA string, it can be re-engineered to recognize a target site based off of a protospacer and a PAM progression . The PAM progression serves as a mediator, ensuring the Cas9 enzyme does not lacerate a necessary fibril of the DNA sequence. Thus, engineers can easily facilitate gene modification by simply editing the composition of the RNA strand conjoined with the Cas9 enzyme. They are not to worry about accidentally fabricating unwanted modifications to the RNA progression because the Cas9 enzyme will not act on it should a PAM not be present. These distinctive aspects of the CRISPR/Cas9 technology promote accuracy by minimizing off-site targeting by the Cas9 enzyme, leaving little to no space for error. Despite the expeditious advances in the field of genetic engineering/modification, bioethical questions are still raised regarding human trials.
Currently, the majority of all genome editing technologies are being tested in the interest of treatment and prevention of human disease. Genome editing shows considerable promise in treating and preventing both single-gene diseases such as muscular dystrophy and sickle cell disease and more complex disorders such as cancers and human immunodeficiency virus (HIV) . A significant amount of research conducted on genome editing directly affects somatic cells (cells that are not egg and sperm cells). However, at the alarming pace that this field of study is evolving, there is no doubt that gene alteration technology to edit germline cells (egg and sperm cells) will be readily available soon, which is where bioethical challenges are raised. The ability to edit germline cells will enable consumers to enhance normal human characteristics, such as height or intelligence. It will also allow consumers to edit their offspring’s appearance .
The second major debacle that is brought into question is human autonomy. When germline editing becomes readily available, it will allow consumers to change normal human attributes in their offspring. This will be a non-consensual act on the parents’ behalf which will impact their offspring. Further, genetic editing will allow consumers to model the perfect human, which in turn, will infringe on a diverse number of societal rights . These societal rights include but are not limited to the right to morality, the right given to doctors to advise patients, the rights scientists are given to conduct research, and the right children should have to consent or not consent to decisions their parents will make that affect their livelihoods.
Based on these ethical challenges, many countries have made it illegal to further study genome editing, more specifically germline and embryo genome editing. However, many more countries are persistent in advancing research and trials into the field.
Genome editing presents an attractive approach and considerable promise to treatment and prevention of human disease. Out of the four common classes of engineered nucleases used in the past decade, CRISPR/Cas9 has shown the most promise regarding accuracy. Its distinct features that make it a favorable tool are its negligible margin of error for targeting off-site DNA progressions via PAMs, its reliance on intertwined Cas9 enzymes and RNA progressions which allow for efficient reprogramming, and its unique PAM system which enables engineers to directly trim an RNA sequence without effectuating complications. However, amidst the steady progress of the genetic engineering field and its universal appeal to researchers, one can make a point to further analyze bioethical inquiries that have arisen before implementing further studies, trials, and inevitably, introducing this technology to the healthcare market.
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Doudna, J. A., & Charpentier, E. (2014). The new frontier of genome engineering with CRISPR-Cas9. Science, 346(6213), 1258096. doi:10.1126/science.1258096*, E. P. (2014, July
Jiang, W., Bikard, D., Cox, D. et al. RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Nat Biotechnol31, 233–239 (2013). https://doi.org/10.1038/nbt.2508
Although cancer is a widespread disease that is vigorously studied and researched, we as humans have not yet developed a cure. Most health officials agree that the best way to stop cancer in a patient would be to try and stop it early on. While detecting cancer early might sound like an easy task it can be difficult. Whether it be the steep price or the sparse locations screenings and tests can be hard. But researchers across the globe have been striving for a better solution. One study by researchers at BioScentDx shows that Dogs can accurately detect early-onset cancer through smell. The researchers used a form of clicker training to teach for dogs to distinguish between normal blood serum and samples from patients with malignant lung cancer. After many trials and tests, the researchers concluded that the dogs had a 96.7 percent accuracy of identifying the lung cancer samples and a 97.5 percent accuracy of identifying the normal samples.
But how do the dogs do this? Researchers from many universities, including Stanford Med and BioScentDx, say it is from the very sensitive smell receptors that K9’s have. Dogs have 10,000 times more sensitive smell receptors than humans which would allow them to smell and identify various more biologic compounds such as cancers. Other researches have said that dogs are able to smell cancer due to the olfactory ability that they have. This allows them to detect very low concentrations of alkanes and aromatic compounds generated by malignant tumors through urine or in the breath of humans. The very thought that dogs could detect cancer dated back to 1989 when it was published in The Lancet Medical Journal.
If the research is deemed accurate by secondary trials and dogs are able to detect cancer, it could be very exciting for the medical community and for cancer research. The company BioScentDx has said that it would pave the way for further research along two paths, both of which could lead to new cancer-detection tools. One is using canine scent detection as a screening method for cancers, and the other would be to determine the reach of K9’s senses. The company plans to use canine sent detection to develop a non-invasive way of screening for cancer that would be less expensive as well as more accessible.
Overall, the use of cancer sniffing dogs in the medical and health science fields could be a huge discovery for the livelihood of many patients. If we already had the K9’s in action we could estimate that nearly ⅛ of all the people with cancer would have been screened earlier, stated the Center for Disease Control, which potentially could have saved hundred of thousands of lives. In the near future, many companies are planning on further testing canines to sniff cancer and other diseases too. And if it works, dogs could change the world for the betterment of society.
Comparative Analysis of Effectiveness of Apoptosis
As there are more than a million cases in the United States annually, and over 500,000 Americans die each year, cancer can be characterized as the continual unregulated rapid increase in the number of abnormal cells. The use of mouse model cancer cells serves an utmost significance in the field of research as the causal link to carcinogenesis, the initiation of cancer formation, and cancer genes can be used to test and develop new therapies. Chemotherapy is the most common approach when looking at cancer treatment, where cancer cells are killed using chemicals that are toxic to living cells. Antioxidants maintain homeostasis and the maintenance of cell integrity in human immune systems and have shown to increase the life quality of the patients. The purpose of this experiment was to determine which antioxidant will be the most effective at different volumes to promote apoptosis in mouse osteoblastic cancer cells (MC3T3). The hypothesis was that as the volume of antioxidant solution increases, then the apoptosis of MC3T3 will increase. The experiment was conducted as MC3T3 cells were first grown for 24 hours, then the antioxidants Lycopene, Resveratrol, and Vitamin C were then added to the cells. The cells were once again grown for 24 hours, and after the 24 hours, cell viability was measured through MTT assay. The results show, the cell viability of Lycopene at 100uL was 27%, it was the most effective in promoting apoptosis. The hypothesis was supported to the extent that Lycopene and Resveratrol were successful in promoting apoptosis at certain volumes. This experiment can be applied to help cancer patients by offering another treatment or in combination with chemotherapy to increase the life quality of the patient.
The purpose of this experiment is to determine which antioxidant will be the most effective at different volumes of the antioxidant, to promote apoptosis in mouse osteoblastic cancer cells.
Independent Variable – The volumes of the different antioxidants that will be induced into the cells
Dependent Variable – The cell viability of each antioxidant at different volumes, which will be measured using MTT assay
Control – Micro-wells not being induced with any treatments
Experimental Control – Etoposide, which is a chemotherapeutic agent proven to promote apoptosis and most prescribed chemotherapeutic agent in the world
Introduction & Background Research
Over 500,000 Americans die of cancer each year, as there are more than a million cases in the United States annually (Cooper, 2000). Cancer can be characterized as the continual unregulated rapid increase in the number of abnormal cells, leading to a group of cancer cells, which is a tumor (Cooper, 2000). Cancer is caused by carcinogens that start the growth of cancer cells; carcinogens are substances that promote or initiate the growth of cancer cells (Cooper, 2000). Cancer cells grow with continual proliferation, rather than healthy cells that respond appropriately to the signals that control normal cell behavior (Cooper, 2000). Tumors are a group of cancer cells that can be categorized into either benign or malignant (Cooper, 2000). A benign tumor is similar to a wart where it does not spread cancer to other tissues and other parts of the body and stays in a central location (Cooper, 2000).
On the other hand, a malignant tumor is capable of spreading to other tissue as well another parts of the body through lymphatic and circulatory systems (Cooper, 2000). Among the various unique types of cancer, they are classified as either sarcoma, carcinoma, lymphomas, or leukemias (Cooper, 2000). Carcinomas, which are the most common, and occurs in 90% of human cancer cases, are malignancies of epithelial cells, which are sheets of cells that line internal organs and cover the surface of the body (Cooper, 2000). Sarcomas are tumors that are solid in connective tissues, like bones, cartilage, and muscle, which rarely occur in humans, and lymphoma or leukemia rarely occurs with 8% human cancer cases, comes from the cells of the immune system and blood-forming cells (Cooper, 2000).
Metastasizing, which is the development of a malignant growth that is secondary to the primary site of cancer, which grows at a distance and spreads to other sites in the body (Macedo, Ladeira, Pinho, Saraiva, Bonito, Pinto, Goncalves, 2017). The third most frequent site of metastasis is the bone, which has a relative incidence percentage of 65-75% (Macedo et al., 2017). The three types of bone metastasis are classified as Osteolytic, Osteoblastic, or mixed (Macedo et al., 2017). Osteoblastic cancer is seen as the breakdown of the new bone similar to that of prostate cancer, small cell lung cancer, carcinoid, and Hodgkin lymphoma (Macedo et al., 2017). Bone metastasis is a sign that the disease has spread to other organs and predicts a short term probability of the disease in cancer patients (Macedo et al., 2017). Once cancer has spread to the bone, the chance of it being cured is low, though treatment can be applied to slow its growth because new drugs and treatments have shown a better quality of life and improved life expectancy (Macedo et al., 2017). Both Filipa Macedo and colleagues and Geoffrey Copper discuss the impact that cancer has left on society. Though they both discuss metastasis and how it leads to malignant tumors, Macedo looks at it only from bone metastasis, while Cooper analyzes metastasis through cancer in general. As they look at cancers through different outlooks, Macedo categorizes the four main types of cancers, and Coopers analyzes Sarcoma cancer, specifically bone cancer and the three types of bone cancers.
When looking at metastasis it is also essential to look at carcinogenesis, which is how cancer formation starts, and neoplastic development, which is the formation of diseases that conditioned to cause tumor growth, which occurs when oxidative stress, damage from cancer cells, go unrepaired, that among humans and animals, research has shown the degree of biological and genetic similarity (Balmain & Harris, 2000). Mice develop tumors in the same tissues and amazingly also in the same histopathological course as humans even though they are much smaller animals with a higher metabolic rate (Balmain & Harris, 200).Humans have similar characteristics such as anatomical, cellular, and molecular to mice, which have the utmost importance in function and critical properties in cancer (Tratar, Horvat, & Cemazar, 2018). The use of mouse models serves the utmost significance in the field of research as the causal link to carcinogenesis and cancer genes can be used to test and develop new therapies (Tratar et al. 2018). Both Balmain and colleagues and Tratar and colleagues discuss how mice cells are similar to human cells are similar biologically to the extent of their higher metabolic rate and other small factors. Tratar goes onto discuss how the use of mice cells can be useful in research to apply applications on to human cells. In contrast, Balmain looks more into the histopathology, examination of tissue at a microscopic level, similarities of the cell, such as the presence of Telomerase, an enzyme that helps maintain chromosome integrity in both human cells and mice cells.
As mice cells serve as useful models of human cancer cells, the use of mice cells can be used to test new treatments such as chemotherapy drugs and antioxidants. Chemotherapy is the most common approach when looking at cancer treatment, where cancer cells are killed using chemicals that are toxic to living cells (Thyagarajan & Sahu, 2018). As chemotherapy has shown to be effective, it has many consequences such as DNA damage and damage to DNA replication, and it induces cell death into living healthy cells (Thyagarajan & Sahu, 2018). As these effects of chemotherapy affect the quality of life of cancer patients, there is a need for alternative treatment. Antioxidants maintain homeostasis and the maintenance of cell integrity in human immune systems (Pham-Huy & Lien Ai et al., 2008). Many different types of antioxidants can be found in vegetables; they have been shown to delay or prevent oxidative stress (Pham-Huy & Lien Ai et al., 2008). Antioxidants also serve as an effective treatment for cancer as studies have shown an increase in apoptosis, programmed cell death, in different cancer cell lines, and inhibit cell proliferation (Pham-Huy & Lien Ai et al., 2008). Both Pham-Huy and colleagues and Thyagaran and Sahu show the effectiveness of antioxidants as an anticancer treatment. Though Thyagaran and Sahu examine the relationship between the effectiveness of chemotherapy and its effects on antioxidants efficacy. Pham-Huy, in contrast to Thyagaran and Sahu, looks at specific antioxidants and their effectiveness in different clinical studies. As there are many different types of antioxidants, such as Lycopene, Resveratrol, and Vitamin C (ascorbic acid), it is crucial to examine their effectiveness against cancer cells and their effectiveness when compared to chemotherapeutic drugs such as Etoposide.
Lycopene, which is mainly derived from tomatoes, tomato products, and other fruits, has shown to be a potent antioxidant and to reduce the incidence of cancer (Johary et al., 2012). Dietary carotenoids, a product of plants, such as Lycopene, have shown to be connected to the decreased risk of cancer and the maintenance of good health (Johary et al., 2012). Recent studies have shown Lycopene as an anticancer agent as its ability to inhibit metastasis, increasing the antioxidant response, preventing oxidative stress, and promoting apoptosis (van Breemen & Pajkovic, 2008). Lycopene has the potential, as a natural product, to be used in cancer treatment to increase the rate of apoptosis based on the studies that show Lycopene supplements when used against prostate cancer have shown reduced risk, anticancer activities, and chemoprevention, chemicals that prevent or slow development of cancer activities. Johary examines Lycopene as a dietary carotenoid and its many properties, which lead to beneficial effects such as preventing cancer. Van Breeman and Pajkovic look at the properties of Lycopene as an antioxidant and evaluates its efficacy by looking at clinical studies. Both authors view Lycopene from a different perspective, though they both establish it to be an effective antioxidant against cancer.
Resveratrol is a product containing high levels of naturally occurring phenols (organic products), which is derived from plant sources, berries, peanuts, and grapes (Bishayee, 2009). Resveratrol has shown to be effective against the three stages of carcinogenesis: initiation, promotion, and progression (Bishayee, 2009) Resveratrol has a robust anticancer property as it can inhibit proliferation in human tumor cells (Ko et al., 2017). As Resveratrol can stop carcinogenesis, it has brought much attention to the prevention of cancer, a therapeutic drug, treatment researchers (Bishayee, 2009). Resveratrol can make cancer cells sensitive to chemotherapeutic agents and reverse multidrug resistance to cancer cells when used with drugs that are used clinically because this shows its efficiency in fighting cancer cells (Bishayee, 2009). Reports have shown Resveratrol can make cancer cells sensitive to chemotherapeutic agents and reverse multidrug resistance to cancer cells when used with drugs that are used clinically. Ko and colleagues examine Resveratrol specifically when used for cancer treatment and look into studies that inhibit Resveratrol into variety of cancers. Bishayee also examines resveratrol when used for cancer treatment, but specifically looks into Rodent studies which can further be implicated. Both Bishayee and Ko and colleagues see resveratrol as an effective anti-cancer treatment as they both discuss the effectiveness even looking at different types of cancers.
Ascorbic acid romote cell proliferation and cell differentiation in cancer cells (Yang & Seo, 2013). Ascorbic acid, also known as Vitamin C, in recent studies, has shown its ability to promote cell proliferation and cell differentiation in cancer cells (Yang & Seo, 2013). Ascorbic acid has also been shown to increase the osteogenic formation of bone in bone cancer cells when treated with cell differentiation (Yang & Seo, 2013). Vitamin C also has been shown to increase the survival rate when inhibited with organisms with cancer cells and increase the survival rate of the mice (Yeom et al., 2009). The administration of ascorbic acid in cancer patients when they were in the terminal stage showed the increase in the quality of life because of this experiment shows the sufficiency of ascorbic acid when induced in sarcoma cells (Yeom et al., 2009).
Etoposide, an epipodophyllotoxin, a substance occurring naturally in Mayapple Plant, is structurally similar to another chemotherapeutic drug vincristine, a chemotherapeutic drug that is in the same group of drugs called alkaloids (Stadtmauer, 1989). Recent studies show the effectiveness of vincristine as an active agent against bone cancer, which is structurally similar to Etoposide (Stadtmauer, 1989). Higher doses of Etoposide will increase the rate of apoptosis in the cancer cells as shown to be an active agent when used at high dosages with other chemotherapeutic agents when applied to bone cancer (Stadtmauer, 1989). Etoposide has been shown to inhibit the growth of pancreatic cancer cells and induce apoptosis (Zhang & Zhang, 2013). Etoposide is a principal chemotherapeutic agent that has been shown to treat many types of cancers because, for more than two years, it remains the most prescribed drug for cancer treatment (Zhang & Zhang, 2013).
Research on antioxidants and their effects on cancer cells have shown that higher dosages increase the rate of apoptosis. Though they all increase apoptosis when the dosage is increased, it is also essential to discuss which treatment would be the most effective in comparison to Etoposide, which is a known chemotherapeutic agent. A study that examines the effects of Lycopene, Resveratrol and Vitamin C (ascorbic acid) on mouse osteoblastic cells (MC3T3) would enable us to see which one is the most effective compared to Etoposide.
As you increase the volume of the antioxidant solution in presence of mouse cancer cells, then the apoptosis of the mouse cancer cells will increase, because studies on antioxidants have shown to increase apoptosis in many different cancer cell lines.
Mouse Osteoblastic Cancer cell line (MC3T3) and other special instruments were provided by the University of Arkansas Little Rock. Throughout the whole experimentation all equipment was provided sterile and goggles, aprons, gloves, and other lab safety precautions were taken. An incubator was used throughout the experiment and was constantly at 37 C with 5% CO2 atmosphere.
To first begin collecting cells from the original medium in the original mammalian cell 1 culture flask, cells were washed with PBS (11.9 mM phosphates, pH 7.4, 13.7 mM 2 NaCl, 2.7 mM KCl). Then using Trypsin-EDTA (0.05% Trypsin/0.53 mM EDTA in 3 HBSS without sodium bicarbonate, calcium and magnesium), cells were dissociated and placed into a incubator for 5 minutes until cells detached. Then a centrifuge at 500 g for 4 5 minutes was used to separate both medium and cells. Cells were then resuspended into a 10 mL medium with 10% FBS which was supplemented with antibiotics penicillin 5 6 (500 units/ml) and streptomycin (500 units/ml). Cell density was then measured by 7 staining cells with Trypan blue dye by using a hemocytometer. Cells were then seeded 8 into a 96 well plate at 1.0 – 1.5 x 8000 cells per well with 100μl medium, where only 24 wells seeded. Cells were then left to be grown in the incubator for 24 hours.
1 A substance used to promote cell growth of microorganisms 2 Polyphosphate Buffer solution, used to maintain pH 3 is used to remove cells from original culture 4 A machine used to separate liquids or solids of different densities 5 Fetal Bovine Serum, used to increase within medium 6 Helps control bacterial contamination
Antioxidant solutions were prepared by either crushing Vitamin C pills or making solutions from gel tablets. 2 determinations of 10μl, 50μl, and 100μl of Vitamin C, Resveratrol, and Lycopene were made. Each antioxidant at each specific volume was placed into 2 wells. Etoposide at 10μl was also placed into wells. 2 wells were left with no treatment to serve as a control. Cells were then left to be grown in the incubator for 24 hours.
The method that was used to measure the cell viability in this experiment was MTT assay. MTT assay is the process by which analyzing the activity of mitochondrial-dehydrogenase as a way to the number of living cells. To first begin, the cells were washed with FBS twice that was medium free of any medium. After they were washed, 10μl of MTT solution (5 mg/ml) was added to the cell culture and then the microwell plate was left in the dark in the incubator for 4 hours because it is light sensitive. After the 4 hours, the medium was removed and the cells were lysed with 9 dimethyl sulfoxide, which helps dissolve MTT formazan which was produced mitochondrial succinate dehydrogenase. The cells were then placed into a microplate cell reader at 570nm to measure the absorbance of the purple color of each well produced by the MTT solution. Determinations were performed in duplicates for each treatment for better statistical analysis. The results of MTT assay were presented as a % showing the control values obtained from untreated cells. The cell viability was calculated using the % of dead cells = (A sample – A blank) /(A control – A blank) x 100.
7 Helps control bacterial contamination 8 Dye used to identify living tissue or cells 9 To undergo Lysis, breaking down of cell membrane
Data / Observations
Absorbance at 570 nm Trial #1
Absorbance at 570 nm Trial #2
10 μL Etoposide
Vitamin C (A)
Vitamin C (B)
Vitamin C (C)
Table 1. Absorbance Readings at two different determination of antioxidants at various volumes
Absorbance at 570 nm
10 μL Etoposide
Vitamin C (A)
Table 2.. Effects of Antioxidants at 10uL on Cancer Viability. Shows the prominence of each antioxidants effect on the cancer cell culture
50μL of Antioxidants
Absorbance at 570 nm
10 μL Etoposide
Vitamin C (B)
Table 3.. Effects of Antioxidants at 50uL on Cancer Viability. Shows the prominence of each antioxidants effect on the cancer cell culture.
100μL of Antioxidants
Absorbance at 570 nm
100 μL Etoposide
Vitamin C (C)
Table 4.. Effects of Antioxidants at 100uL on Cancer Viability. Shows the prominence of each antioxidants effect on the cancer cell culture.
Graph 1. Viability of Cancer Cells in 10uL of Antioxidant Solutions. Displays the different antioxidants’ effects on the cancer cell culture
Graph 2. Viability of Cancer Cells in 50uL of Antioxidant Solutions. Displays the different antioxidants’ effects on the cancer cell culture
Graph 3. Viability of Cancer Cells in 100uL of Antioxidant Solutions. Displays the different antioxidants’ effects on the cancer cell culture
Table 4. ANOVA Test Results. Shows the statistical variance of data and since the f-value is lower than the p-value in this case, the data is not statistically varied. Since the p-value is higher than 0.05 (> 0.05) it is not statistically significant.
Figure 1. MC3T3 cells after being detached and added to medium
Figure 2. 96-well plate. Contains only the cancer cells and antioxidants before growth
The expected result of apoptosis increasing when increased volumes of antioxidant the solution was true to a minimal extent. The results of some of the antioxidants at specific volumes had the opposite effect, where the cell viability of MC3T3 increased significantly. The cell viability of Vitamin C among the three different volumes, increased 243%, 298%, and 868% respectively. This result could be due to the fact that at a certain point of volume of the antioxidant solution, the cells will then feed on the Vitamin C increasing the viability significantly. Fernandes and colleagues looked at Vitamin C at different volumes and saw a similar result that at cell viability, was increased in cells treated with low volume of Vitamin C with respect to untreated bone cancer cells. Etoposide did not increase apoptosis significantly though 89% of cell viability was present after 10μL was seeded. Some antioxidants at specific volume were more effective in promoting apoptosis than the Etoposide. Lycopene at 100μL was the most effective in promoting apoptosis as cell viability was 27%, though at volume of 10μL and 50μL, it was 136% and 1162%. .The same effect that was on Vitamin C can be seen on Resveratrol as at a certain volume, the volume of the antioxidant will feed the cells until a certain volume has reached. For resveratrol, the opposite effect can be seen as at a lower volume of 10μL the cell viability was the lowest and as volume was increased the cell viability exponentially An ANOVA test was performed, and it ended with a p-value of 0.58602 and an f-value of .55892. This signifies that the data has an extreme variance between each treatment since they all had different results. Since the p-value is above 0.05, we have failed to reject the null hypothesis which is that there is no correlation between antioxidant treatment and cell viability. Within the experiment, there may have been some possible error as the vitamin C solution turned the well yellow while every other well had a more purple color. This may have been related to the vitamin’s natural color, but it could have influenced the results of the experiment. Due to time constraints, only 2 determinations were performed. Contamination could have occured as cells were being grown in an incubator with other organisms, or light contamination as MTT assay has to be done in a dark environment.
The hypothesis that as you increase the volume of the antioxidant solution in presence of mouse cancer cells, then the apoptosis of the mouse cancer cells will increase, was supported to the extent at which Resveratrol and Lycopene increased apoptosis when induced. Each antioxidant significantly demoted growth to a certain extent within the cancer cell apart from the Vitamin C solution. Furthermore, the polychemotherapy can be investigated by looking at not only chemotherapy but as well as a combined treatment of antioxidants and chemotherapy. Research can also be done on looking at the patient’s quality of life as they are being given normal chemotherapy and to also see if it is different as antioxidants are introduced.
This project required an immense amount of outside help from Dr. Nawab Ali and Mrs. Puja. The supervision of a qualified scientist of her expertise was necessary in order to follow procedures accordingly. Dr. Ali also gave great advice throughout the process. Without the assets and resources provided by the Dr. Ali’s lab, there would have been no way for this project to come about. Mrs. Becker and Mrs. Norris contributed a significant amount of help throughout the research and statistical analysis process.
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