Results

The second to the last week was surprisingly exciting. I was thrilled to see if we could find any associations using the lab data. Dr. Allison and Melanie assisted me in using statistical approaches to analyze my data. One of the interesting things that we found was that child IQ was predicted by genotype at one VDR SNP (RS7975128, p=0.01), and maternal smoking within the child’s first 7 years of life (p=0.03). However, the interaction between the VDR SNP and maternal smoking was not significant (p=0.40). The A/G and G/G genotypes of RS7975128 were associated with lower IQ, as was the occurrence of maternal smoking. I also attended a lab meeting and a clinical meeting.

BuSy Week With the ADHD samples!!!!

On Monday, I was able to plate my ADHD DNA using the Biomek robot. This made me extremely excited because I was finally able to start on my project. I left the plates to dry overnight at room temperature. The following day, I began working on my assays involving the ALAD gene and the VDR gene. I made 18 Taqman plates for 9 assays and ran 14 of those plates onto the PCR machine. I then scanned them on the 7900HT machine. Most of my TAqman went well except for two plates. Next, for Wednesday, I had to re-run the two plates that failed. I also ran 6 more plates on the PCR machine and made 2 plates for a single assay. I scanned those plates as well and they turned out great. For the rest of the day, I had to plate 20 extra plates using the Biomek robot. On Thursday, I made 5 Taqman plates in the morning. I ran them using the PCR machine and scanned them afterwards. I then added markers and detectors. Fortunately, they looked great. In the afternoon, I ran about 5 more plates and scanned them. In addition, the fire alarm went off again for the second time since I have been in CHG. On Friday, I really enjoyed the talk by Ms. Cleaver. It gave me many tips in applying for college. After that, I went to my lab and ran and scanned some plates that I had made prior. I also helped Kaia by making two agarose gels for her and running her PCR samples. Some looked good but the rest failed. Chris then helped me submit my data so that I could view them with a statistician next week. Unfortunately, two failed to submit but I’ll be able to do that on Monday. Overall, I had an exciting and this week was probably the busiest for me by far.

FAmily Visit

I was able to observe my first family visits on a Friday. So far, that was probably one of the highlights of my experience with the program. I watched as the entire family of four completed a series of behavioral and psychological tests as well as developmental histories to see if they could be diagnosed with ADHD. There are three rooms in the clinic area: a computer room, an interview room, and a blood room. One member of the family goes into the computer and interview room at a time. In the computer room, letters appear on the computer simultaneously at varying time intervals and every time a letter, other than “X”, appears, the participant has to push the space bar. This test takes place for exactly fourteen minutes. After a while, one of the young participants that I watched became bored of the test, so he continuously and randomly just began pushing the space bar, increasing his impulsitivity score. The interview room is where the psychologist assesses them and gives them an IQ test by asking several questions as well as by allowing them to complete spatial and hands-on tasks. Finally, after everyone in the family completes these tests, there blood is drawn and set to the DNA bank. Unfortunately, one of the children refused to get a blood draw. Overall, the entire family visit lasted for a period of three hours and thirty minutes.

ADHD

On Monday, I continued working on my methods section until Chris, my mentor, arrived. Surprisingly, one of my friends from school was assigned to the same lab as me and she had just begun working there. So, she had to observe Chris and I for that day. Prior to lunch, I worked on two PCR plates and four Taqman plates. Then after lunch, I had to make an agarose gel to test my PCR. My PCR worked; however, it was not that great since some of the wells looked as if they were missing DNA samples. I assisted my friend in practicing using the pipette. Then, I completed one more PCR plate and scanned my Taqman plates afterwards. Fortunately, all of my Taqman worked out well. For the rest of the time, I worked on my methods section again and completed my blog.

Tuesday, I worked on a Taqman plate and two PCR plates. After running them on the machine, I had to wait until they finished, so that they could be tested on an agarose gel. After lunch, I tested the samples on the gel. My results were better than my results on the previous day. So, I completed two more PCR plates using a different primer, and ran them on the PCR machine. Then Chris taught me a new method using Sephadex to filter the DNA from our PCR. It was fun being able to learn a new method. This was a gel like substance that allows the DNA to separate from the excess materials used during the PCR reaction. After that, I worked on my blog.

I had a wonderful Independence Day celebration with my friends on Wednesday and returned to work on Thursday. I felt so excited to receive my first paycheck. When I came to the lab, I made an agarose gel and ran my previous PCR samples on it. It actually worked out well. Afterwards, I made another PCR plate prior to lunch. I made another PCR plate after lunch and I also began labeling my ADHD plates. I then waited for my PCR plates to finish running so that they could be tested on an agarose gel.

Friday was a great day! I began my day by making a PCR plate. I then ran my PCR agarose gel that I made earlier in the week. Fortunately, all my samples worked. Next, I ate lunch early because I was invited to observe a family visit. This is when the proband as well as the entire family come into the clinic area to be tested for ADHD.

Taqman and PCR

Monday was quite interesting. A couple of minutes upon arriving at the Center for Human Genetics Building, the fire alarm went off and everyone had to be evacuated outside the building. When we came back in, I looked at the results for the last for Taqman plates. Sadly, two of the plates failed to amplify, which means that I have to re-do these plates. Then, Chris showed me how to do PCR. He explained the functions of the various ingredients in the master mix. For instance, the taq is an enzyme used for replication and amplification of a DNA fragment. He also showed me how to work the PCR machines. When I finished lunch, I had to re-do the plates that failed. Chris then allowed me to do PCR by myself, which was so exciting. For the remainder of the day, I analyzed the rest of the Taqman plates. In addition, I went into Pub Med and searched for the gene ALAD and lead and neurotoxicity.
Tuesday was a fun day also. I enjoyed the Primate Center tour. I have never seen lemurs in the past and it was amazing to see how closely they resembled humans. In the lab, Chris and I went to use the Biomek robot for transferring DNA into the 384 deep-well plates. At the same time, Robin came to my lab for a lab visit. We talked about various aspects of my project as well as the poster. After lunch, I had a meeting with Dr. Allison and Melanie. We looked at different data and statistics regarding smoking, genotypes, and polymorphisms in relation with ADHD. Amazingly, we found many significance in the data. We also discussed the status of the materials for my project. I then headed back to the lab and did two PCR plates. We actually had a small going away party for two members of the lab who are going to be relocating to Miami. We had some cake and everyone in my lab joined in. Overall, Tuesday went well.
On Wednesday, I was able to make an agarose gel. It was somewhat nerve wrecking to handle even just a small amount of ethidium bromide since it could possibly affect my future children’s DNA. It was, however, interesting how we made a gel with multiple rows of wells rather than just one. The purpose of the gel was to see if our PCRs worked. After running the gel, we took a picture of it and measured the band sizes. Fortunately my first PCRs were great. After lunch, I completed one more PCR plate, and then I proceeded to do two Taqman plates for different SNPs. I then started working on my blogs for thirty minutes. Afterwards, Chris and Dr. Allsion helped me out with my introduction.
On Thursday, I met with Dr. Allison in the morning. She informed me about the upcoming family visit for the ADHD research study patients, wherein they go through a series of tests and get their blood drawn. I am very thrilled to see that. After that, I went to my lab and finished two PCR plates. When lunch was over, I ran the samples from the plates onto the agarose gel to see if my PCR was successful. For the remainder of the day, I completed some more Taqman for patients enrolled in the hostility study.
Finally on Friday, I started planning out the methods section of my poster. Also, I searched about the various SNPs in the genes that I will be studying. I had to find its role and its functions. I made three PCR plates using different primers and ran them on the PCR machine. After that, I made an agarose gel to test my plates on. When lunch was over, I tested my samples from my PCR plates. As it turned out, my PCR turned out well. For the rest of the day, I worked on some more PCR plates and headed to the Schiciano Auditorium for the seminar speaker !! This week was a busy week, but it turned out great!

Attention-Deficit/ Hyperactivity Disorder

Hello everyone! To formally introduce myself, my name is Alexa Nicole Celerian. I am a rising junior from Josephine Dobbs Clement Early College High School. It is an amazing school located at the heart of North Carolina Central University. I was born in the Philippine Islands, and I resided there for more than ten years. Throughout my childhood years, I learned how to speak four different languages. I enjoy sports; however, my most favorite is swimming. I have a family of four consisting of my mother, father, and a 14-year-old brother. My family is my inspiration. In the near future, I aspire of becoming a successful cardiologist. As a child, I have always dreamt of becoming a medical doctor. I love working with people, especially children; so, I might even consider the field of pediatric cardiology. I feel that the Howard Hughes Program will provide me with a greater body of knowledge.
My first week in the program was extremely interesting. There was so much to learn and I have never been exposed to so many lab instruments/ technologies in the past. On Monday, my Principal Investigator, Dr. Allison Ashley-Koch, cordially greeted me. We then walked to the Center of Human Genetics, the area wherein I will be primarily doing my research. During my first day, I had to sign a Confidentiality form, and then Dr. Allison began introducing me to my project. It involves trying to find out if environmental factors such as smoking, lead exposure, alcohol use, and so forth, heightens the risk or is related to Attention- Deficit/ Hyperactivity Disorder (AD/HD). I read about the basics of Genetics in medicine to gain a better understanding of the subject. In addition, I read more about assessing AD/HD. I learned that AD/HD involves hyperactive-impulsive or inattentive symptoms that caused impairment and were present before seven years of age. Overall, it was a day of trying to understand the concept of the main subject I was trying to research.
Tuesday was another exciting day! It mostly dealt with the clinical side of research. After meeting with Dr. Allison, I was introduced to Linda Exelbierd, who is the AD/HD study coordinator. She is in-charge with consenting the patients and informing them of the benefits/ risks of their study participation. The proband is usually a child under the age of twelve who has AD/HD. She tries to consent as many of the proband’s family member to see the history of AD/HD within that family. To participate in the study, the family often has to go through many steps. They have to answer many questionnaires, developmental histories, and general information. In addition, they have to be checked for their attention span as well as have their blood samples are drawn. The process then repeats after two years. Linda actually showed me several pedigrees of patients she inputted in the computer. It was interesting to see the various generations affected with AD/HD. After meeting with Linda, I was introduced to Shelley Keats, another clinical employee. She inputs questionnaire data into the computer and she is also present during family meetings. As a matter of fact, I may be able to observe a family meeting when they have one! One very striking thing that we talked about was a disorder known as Trichotillomania, wherein people pull their hairs or pick their skin excessively to the point of baldness. According to Shelley, patients whom she has spoken to in the past told her that they do not feel any pain when they pull their hairs or pick their skin. Tuesday was very intriguing.
Next came Wednesday. The highlight of that day was surely getting an I.D. badge. It made me feel “Official” !! After that, Melanie Kail and a fellow personnel showed me how they put patient data into the computer. Melanie will help me organize my data after working in the lab. We looked through the data and queried the factors that might affect AD/HD. We analyzed the results and placed it in a spreadsheet. Unfortunately, we didn’t see much correlation but it might have just been due to the fact that we did not have that big of a population size. One significant thing though is that all the children whose mothers drank while pregnant were affected. I was asked to observe a clinical meeting as well. They were mostly talking about regulations dealing with accessing patient files. Then, for the rest of the day, I completed several IRB modules as well as Online Safety Training Sessions.
On Thursday, I met with Dr. Allison again in the morning and she and I talked about my first week as well as the activities that I participated in during the morning with the Howard Hughes Program. We read about the role of dopamine transporter (DAT) genotype and maternal prenatal smoking on children with AD/HD. Dopamine transporter is the system that helps neurons with transporting messages/ impulses. The journal stated that children with two copies of the DAT allele and had prenatal smoke exposure had greater hyperactive-impulsive scores compared to its contrary. For the rest of the day, I completed some more safety training and I read more excerpts from journals.
Lastly, Friday was perhaps the best part of my week. Dr. Allison was teaching me about the structure of a DNA. I then attended a session with Julie Rochelle as well as other new personnel in the Center for Human Genetics. She discussed lab safety rules, the dress code, and other ethics rules. Next, I was able to go to the lab and start practicing using the pipet with my mentor Chris. He gave me a tour of the lab and I was very fascinated by the various gadgets that they use. I have never seen such things in my life. He explained to me how the TaqMan works. It is very similar to a PCR except it involves probes that emit light waves. He explained how I would be looking at various DNA’s from patients with AD/HD, attempting to look for a single nucleotide polymorphism mutation that is common with most/ if not all of them. So, I am highly invigorated to start my research. I cannot wait to see what more will come from the next several weeks!