Alejandro Rios' Summer

Week 5

2005-07-14T11:40:00.000-05:00

We prepared ourselves for the p450 experiment by making 1 mol of KPO4, NADPH, G6PO4, G6PDH, and a 1:4 dilution of MgCl. Then we made our tubes in preparation for the experiment. We then set up charts of ‘time dependent’ and ‘protein dependent’ experiments. Today we proceeded to complete the experiment doing the time dependent after a lecture of the procedure. We only had time to run the timed experiment, so Dr. Clarke ran the protein experiment over night. We did a binding experiment with Dr. Harp. We talked about IC50 and how we were going to set up the experiment. Then we talked about how to set up the experiment with the tubes and the different concentrations of drug. Since we were not able to finish our own drugs, we used some that Dr. Harp already had for us. Then we proceeded to interpret the data from Dr. Harp’s experiment since we did not finish. Today Dr. Clarke told us that we were to do another experiment like the one from Monday and Tuesday, but on a smaller scale. We will be using different concentrations of DPP and of our individual drugs (Phenacetin, Dextromethorphan, ibuprofen, and Terpenadine). This experiment will tell us if DPP is an inhibiter of the drugs.

Week 4 with Dr. Harp

2005-07-11T08:51:00.000-05:00

We preformed two tlc’s to determine our product but no such luck. So we had to use a column chromatography to determine if we had any product in out solution. Column Chromatography is just like tlc just that the produce travels downward due to gravity. Using tubes we collected samples of the solution to run tlc’s on the individual tubes. Today we flushed our column chromatography solution and rotervaped our completed DFB and prepared an NMR test solution. We learned about how we use an NMR to determine the products magnetic field and from that we can see what is in our solution. We went to wake forest to se a 300 mhz magnet that was used for the NMR. Then we went back to PTCRC to heir a lecture that Dr. Harp had to give. Today we helped clean the lab up and organize it a little. Then we proceeded to rotervap our flush and tube solution from the other day. Once completed, we prepared solutions for the rat dissection. Dr. Harp showed us how to kill the rat properly and the important parts of the dissection.

Week 3 with Dr. Clarke and Harp

2005-07-04T14:01:00.000-05:00

We ran the Bradford protein assay again because the results were not reliable. Then we used the Genios machine to test the concentration of the enzymes. Last we compared and analyzed our data with one another. The nex day we helped Dr. Harp with children from a summer program making aspirin using salicylic acid and phosphoric acid as well. Then heated and filtered using filter paper. It was fun working with those kids. We helped Dr. Harp with the same kids,the next day, to separate the caffeine from tea. First we made the tea. Then we used dichloromethane and sodium hydroxide and magnesium sulfate to separate the caffeine and water from the tea. Then they scraped the caffeine and put it in to some vials and took them home. I've never work with kids that we that young and so many of them at the same time. With Doctor Clarke, we talked about Terfenadine, Dextromethorphan, S-ibuprofen, and Phenacetin. After making enough of each chemical for everyone, we picked who would do which and we poured some into vials. The amounts varied because we needed one mole of each and that is dependent on the molecular weight. Then we ran out of time so we stopped there to pick up at a later time.

Week 2 with Dr. Harp

2005-07-04T13:51:00.000-05:00

Dr. Harp talked to us about the dopamine transporter and how cocaine and meth affect the flow of dopamine, which causes the high. We Learned how it is necessary to keep a good note book. We worked with DFB and NaBH4 and did thin layer chromatography to distinguish polarity. Then Dr. Harp told us about the distillation and purification technique, which we used with hydrocarbons and later used a vacuum filtration methed, and used theoretical yield to estimate results. Then we set up a found bottom flask with a y-connection, condenser and dryer, all on top of a heat tray. We poured amino alcohol, potassium carbonate, and DMF into the apparatus. We had to start over with one of our solutions due to a small mishap, but we pulled through. Later we learned about nucleophilic substitution-type 2. We did a bunch of tlc's and performed a practice experiment for a class next Wednesday and Thursday. Finally worked on remaking the solution from the mishap from the other day.

Week 1 with Dr. Clarke

2005-07-04T13:24:00.000-05:00

Once at WSSU, the team was given the assignment to look up Diphenylpyraline(DPP). We found it's molecular weight(317.15), found out it was a base, found it's chemical structure and weather or not it was safe to work with(it was). Then we calculated our stock standard solutions and our concentrations solutions. Then we hamoginized some lung, liver and kidney samples and made our solutions out of the rat samples. After completing our calibration samples and our dilutions of the proteins, we performed a Bradford Protein Assay with the Tecan Genios Plate Reader. We then made a calibration curve and analyzed it for accuracy.

Intro to what's to come

2005-07-04T12:30:00.000-05:00

The begining of the week Dr. Harp and Clarke introduced us to the what, where and how of the project. Dr. Harp gave us the info on piperidines, the molecule we will be working with, some of the time. She also spoke to us about the dopimine transporter and how Cocaine and Meth disrupt the flow of dopimine and cause the euphoria that is the known effect of cocaine and meth. Dr. Clarke reviewed calibration curves and two procedures that we will use in the future:Hermaginization(grind) and Differncial Centrifugation(spinning). Both doctors went over the schedule for the next six weeks; it looks like I'm going to be in for a busy few weeks.

The Team

2005-07-04T12:04:00.000-05:00

While everyone else works alone, I have the privilege to work with three other three other people and two doctors. I usually work with Richard. He and I work well together. My other two partners in the lab are Briana and Asia. They usually work together but we all work on the same project. The two doctors that we work under are Dr. Clarke and Dr. Harp. We also go back and forth between labs. The whole team is going to be working hard to make a possible breakthrough that will change the world. I will make sure that I bring you up to date with what's happening in the lab.

Welcome to C.E.R.T.L.

2005-06-24T08:16:00.000-05:00

This summer is going to be an interesting summer, to say the least. From what they tell me(they being the administrators of the program) that I am in for a new experience. For this reason I have dedicated some time out of my now very busy life to bring everyone up to speed on how things are going for me and some quality commentary from yours truly, Alejandro