Week 5 - Natalie

In research this week, I performed the reverse transcription step (turning RNA into cDNA) for the RT-PCR protocol. I did this on E10 Mouse embryo RNA that was purchased, rather than on samples from the mouse fracture study, since all of that RNA has been converted. This E10 mouse embryo cDNA will be used for quality control plates in the future, in order to test the primers of interest for the mouse fracture study. This analysis of the primers will be my main contribution to the study for the summer. To begin this process, I ran a quality control plate for the SFRP1 primer. This plate is used to determine the optimal annealing temperature for the primer. The 96-well plate is set up so that each column is subjected to a different temperature, ranging from 50C to 60C, and each row has a different concentration of the E10 cDNA sample. The goal is to find the temperature where the efficiency is highest at all concentrations. This will then be the temperature used for this primer when the PCR on the actual samples is performed.

I have seen operations in the OR and have gone to the clinic to see patients but I had not yet experienced rounds on patients after surgery. Since the adult reconstruction and joint replacement service is quite busy, one of the residents suggested I shadow in the pediatric orthopedic service instead. I went early one morning to round on two children who had undergone orthopedic procedures. The first patient was an eight year old boy who had a Achilles tendon lengthening procedure. This was performed because the boy was walking on his toes due to the shortness of his Achilles tendon. In this procedure, the tendon is cut in a “Z” pattern, the two pieces are slid relative to one another, and then they are sutured together in the overlapping region. The patient was doing well, although had used his self-administered pain killers a lot during the night. His mother was with him, and was concerned about his pain, but the doctor reassured her that this was normal. The second patient was a fourteen year old girl who had an ACL reconstruction. She was with her mother as well, and was recovering very well, with very little pain. Each visit with the patients was only about 5-10 minutes long, which was a little surprising to me. It was still good to see this other aspect of the clinical experience.

I also attended grand rounds for the adult reconstruction and joint replacement service, as usual. The speaker this week was Dr. Carl Imhauser, from the biomechanics research group. He discussed the planning and early results of an upcoming study on the kinematics of unicondylar knee implants. This study will examine cadaveric human knees on a six degree of freedom mechanical testing robot in the intact case, and then with a unicondylar knee implanted. This study was a great example of a collaborative study between engineers and surgeons. Dr. Imhauser was working closely with Dr. Westrich, an orthopedic surgeon who will perform all of the cadaver surgeries in order for this study to be performed.

Next week will involve more quality control plates for other primers, and hopefully shadowing at New York Presbyterian.