Avoiding Synthetic Drugs to Treat Alzheimer’s Disease: How Effective Is This Solution?
Background: What is Alzheimer’s Disease Like?
Many of you might be familiar with Alzheimer’s disease; maybe a grandparent of yours has it or you’ve heard this condition mentioned at least once. But, what exactly is Alzheimer’s?
Alzheimer’s disease is a neurological condition that’s characterized by memory loss, confusion, disorientation, and a variety of other degenerative symptoms. Individuals with AD have difficulty remembering information and often see themselves as unable to recall family members and even spouses. This is caused by a loss of neurons in the brain, which typically reduces cognitive function. However, how exactly are these neurons lost, and what’s causing them to “die-off?”
According to the Center for Disease Control (CDC), Alzheimer’s was the 6th leading cause of death in 2017 and was responsible for 116,103 deaths.
There are two issues that arise in individuals with Alzheimer’s. The first is senile plaques, which are comprised of a dysfunctional protein called amyloid-beta. Healthy neurons have a protein called an amyloid precursor protein (APP) in their cell membrane; although its function is unknown, there is still significant research being done on this topic. Like many other proteins, APP has to be dissolved and reduced once it’s no longer functional, and this process is done by an enzyme called gamma-secretase. Essentially, gamma-secretase divides the APP into two sections such that the resulting oligomer is dissolvable.
In the case of Alzheimer’s patients, an unknown mutation/alteration causes another enzyme called beta-secretase (hence the name “amyloid beta”) to improperly divide the APP, resulting in an oligomer that’s insoluble. This chain of amino acids, commonly called amyloid-beta, is also chemically sticky and can bind to more chains of itself, resulting in an excess of oligomers bound together outside of a neuron. These plaques tend to stick between the synapses of different neurons and thus affect signal transportation, resulting in declined cognitive function.
The second issue that leads to Alzheimer’s is neurofibrillary tangles, which develop internally. Inside a neuron, there are a series of interconnected “railways” called microtubules, which are held responsible for the transportation of substances throughout a neuron. This “exoskeleton” is stabilized (in other words, “held together”) by a protein called tau, which reacts with the tubulin that comprises microtubules.
In patients with Alzheimer’s, the tau protein is aggressively reacted with enzymes called kinases, which attach a phosphate group to the tau and thus cause it to fold improperly. The tau is no longer able to bind with tubulin and will detach itself from the microtubule, preventing substances from being transported throughout the neuron. Additionally, the hyperphosphorylated tau protein begins to collect inside the neuron and form a neurofibrillary tangle, which also results in many of AD’s typical symptoms.
Treatment Options
The majority of research concerning Alzheimer’s disease is geared towards the development of synthetic drugs. Medication for AD includes Memantine, Rivastigmine, Donepezil, and several other options.
Memantine, which is commonly used to treat mild dementia, has brought slight benefits by permitting clearer thinking and being able to vaguely remember some notions. Like a majority of other synthetic drugs, however, it comes with a multitude of harmful side effects. These include dizziness, constipation, shortness of breath, and even other conditions such as hepatitis and stroke.
Observing the multitude of negative side effects associated with these synthetic treatments convinced many scientists to look for an alternative plan for treating these neurological conditions. A plethora of research concerning the usage of natural extracts in mitigating the progression of Alzheimer’s disease was later published over several years. The variety of compounds were not only less expensive to generate and use, but it also resulted in fewer and less severe side effects. This ended up inspiring me to conduct my own research and discover if I could develop another “natural” treatment plan for patients with Alzheimer’s disease.
How Did I Work With Natural Extracts?
Since oxidative damage and neuro-inflammation are the key pathways implicated in the pathogenesis of Alzheimer’s, I looked towards finding extracts with anti-oxidative and anti-inflammatory properties. Through reading the vast libraries of articles concerning this topic, I noticed that researchers moved towards using one extract/compound in their attempt to slow the progression of AD. I then developed an obscure thought, wondering: can several extracts combined effectively treat Alzheimer’s? I decided to test this hypothesis by looking for natural herbs with the most potent anti-oxidative and anti-inflammatory properties.
I researched and purchased 30 natural extracts, each with a unique set of chemical properties/compounds. To test these extracts, however, I needed an organism that had a similar genetic structure compared to humans and could reproduce spontaneously. I decided to test these natural extracts on the Drosophila melanogaster, otherwise known as the common fruit fly. Drosophila has been used in various genetic experiments over the past century, and I believed it would be the most ideal candidate to observe the effects of these natural extracts.
The prevalent issue was being able to obtain Drosophila that expresses symptoms of Alzheimer’s disease, which led me to use the GAL4-UAS system that would produce fruit flies expressing the Aß42 transgene (which is responsible for generating AD-like symptoms).
What is the GAL4-UAS System?
The GAL4-UAS system is a type of binary expression system that allows a specific gene to be expressed in a particular tissue type (this is why the GAL4-UAS system is referred to as “tissue-specific”).
There are two lines that must be bred for this system to function. The first is a GAL4 line, which contains the GAL4 protein derived from yeast. The other line contains an upstream activation sequence (UAS) which neighbors a reporter gene (this is the target gene that is meant to be expressed in offspring). When the two lines are crossed, the GAL4 protein attaches to the activation sequence and triggers the expression of the reporter gene, which was the Aß42 transgene for my research.
Developing the Extracts
While researching on how scientists utilized these extracts, I noticed that many articles involved mixing the fine powder with deionized water and cooking the extracts at a specific temperature and time. This was largely due to the proteins in these herbs being unaffected and potentially enhanced further. Therefore, I decided to execute this idea with the herbs I had purchased and mixed it with a cornmeal-molasses-yeast medium meant for Drosophila to consume.
With the large number of extracts I had present, I realized that combining all of them might be too difficult as I would have to establish the most precise ratios between all of them. This led me to divide my experiments into Phase I (working with the 30 extracts originally purchased) and Phase II (working with the 12 most effective extracts out of the 30).
Phase I: What Were the Results?
After observing the lifespan of Drosophila upon consuming the 30 different extracts over a period of 60 days, I used Kaplan-Meier lifespan plots to identify the extracts that improved the fruit fly’s longevity the most (this is called a longevity assay).
Now that I was able to determine the 12 extracts that extend Drosophila’s lifespan the longest, I was able to move on to phase II and determine what proportions I had to mix them in to develop the most effective treatment.
What Are Kaplan-Meier Lifespan Plots?
Kaplan-Meier plots are a type of non-parametric (where data does not come from prescribed models) statistical analysis where the lifespan of a particular group is examined after an event X is meant to occur. In the case of my research, that event was the death of Drosophila after consuming the natural extracts.
Phase II And How It Went Down
During Phase II, I developed 5 different extracts with varying proportions and “cooked” them in the same manner as Phase I. Labeling these extracts as the “ALDC Series” (which is short for “Alzheimer’s Disease Control), these extracts were fed to another set of Drosophila that expressed the Aß42 transgene and I performed 3 assays with the experimental flies. During Phase II, I also incorporated a positive (fruit flies that expressed the Aß42 gene and were fed regular cornmeal) and negative (fruit flies that did not express the Aß42 gene and were fed regular cornmeal) control group to measure the efficacy of this solution.
The Lifespan Assay
The lifespan assay I conducted in Phase II was similar to the one occurring in Phase I, where I observed each group of Drosophila on their diet group for 60 days. Based on my results, I was able to conclude that the 4th ALDC variation (also called ALDC4) had Drosophila with the longest lifespan and thus performed the best in this assay.
The Climbing Assay
During the climbing assay, I used a graduated cylinder and made inch markings until roughly 7 inches. I then took the flies from each diet group, placed them in a graduated cylinder, and observed how many were able to climb past the 7-inch mark in 2 minutes. Based on the results, I could conclusively determine that ALDC4 had the highest performance and appeared to increase motor skills most significantly.
The Single Short-Term Olfactory Assay
In this olfactory assay, I took 20 flies raised on each food medium and trained to associate an odor with a 65V electric shock. I used a T-maze (which I created) where the flies are first exposed to the odor Valerian Root while receiving a shock for 1.25 seconds followed by a brief “resting time” for 3.75 seconds. After repeating this cycle for around 60 seconds, I moved the elevator part of my T-maze (the part that is responsible for transporting the flies between each floor of the maze) to the middle range, where there are exposed to Geranium Essential Oil without any shock for 60 seconds. Finally, I moved the Drosophila down to a “choice point” where they were exposed to both odors without any electric shock for about 2 minutes.
The goal of this exercise is to test Drosophila’s associative memory by expecting them to select the odor that was not associated with an electric shock. If the experimental flies were able to demonstrate higher perception and better associative memory, it means the extract was able to delay the progression of AD.
Based on the performance index of the flies, ALDC4 had (once again) proven to heighten Drosophila’s associative memory the most.
Okay, But Why is This Important?
I know what you’re thinking. Great, good for you, but why exactly is this necessary?
Well, with so many synthetic drugs being produced, a majority of them tend to have negative side effects. I believe the risk of taking these manufactured drugs can be mitigated by including natural extracts in treatment for neurological conditions, and this is something that the pharmaceutical industry seems to overlook at times. However, with my own and an enormous amount of other research, larger companies might consider that natural extracts might be a more effective solution for any disease (it doesn’t just have to be neurological).
Key Takeaways:
- Alzheimer’s disease is a condition characterized by memory loss, disorientation, motor dysfunctionality, and a multitude of other symptoms.
- Synthetic drugs have long been a potential solution to treating AD, but the numerous risky side effects led many researchers to consider using natural extracts to slow the progression of this disease.
- I decided to conduct my own research and prove the hypothesis that not one, but several extracts can be combined in specific proportions to more effectively delay the progression of Alzheimer’s.
- This research is necessary to provide a more natural therapeutic solution to neurological conditions like AD, which are extremely overlooked in the pharmaceutical industry.
