Let's not lose focus

We had lab meeting yesterday. We usually hold it in one of the conference rooms over here near our lab (we have two of them on our floor) but since our boss was in between surgeries, we went over to the departmental offices to have our meeting. Now, keep in mind that our lab is as far away from our department as possible without having to go outside. I actually counted the steps one day - it's 1008. That is over 0.5 miles. On days that we have to make several trips to the department, we semi-jokingly tell our boss that he needs to spring for a Segway (sp?). Anyway, I was the only native English speaker in the group and somehow we got on the subject of farms. I spent half of our trip to the department explaining the difference between a ranch and a farm as well as the types of animals on ranches and farms. It was somewhat comical. The two Chinese women giggled when I told them a male pig is called a boar. I am not sure what was so funny about that.

When my friends and I get together, we sometimes discuss the merits of having a PhD for a boss vs. an MD for a boss. I personally prefer an MD. Usually they are in the lab less than a PhD and the good ones know that they aren't science's gift to research. That is sometimes detrimental also. I certainly don't know everything and it is nice to have someone to turn to with questions. I have a very intelligent boss but he doesn't have the bench experience that a PhD would have. What he does have, however, is the ability to remind us what we are doing. So many researchers get so tied up in their one protein or their one gene that they fail to see the patient. Yesterday, during lab meeting, Marlan was presenting a journal article about menigiomas. Meninges are the membranes that line our brain and spinal cord. A menigioma is a cancer that can be quite aggressive although not metastatic. By aggressive I mean that a menigioma will eat its way through bone and tissue. As we were talking about Merlin (a tumor suppressor protein that is missing in menigiomas and schwannomas) Marlan pulled up some of MRIs of a couple of his patients that have menigiomas. One of them was clearly inoperable due to its placement and widespread location. This patient will die. It really helped me to re-focus my efforts and remind me what I am doing. I know that finding a "cure" for this type of cancer (or any type of cancer)will not be a Eureka moment but rather a series of discoveries that will eventually help us to tailor treatments to individuals. So, it is time to get back to teasing apart the molecular pathways these tumor cells use to survive and senesce.

This is my favorite time of day at work. I try to get in right around 8 am because nobody else is here. It is quiet and I can get my work started. We just had another snow storm last night and I have a great view to the north. Everything is frosted white and with the sun out this morning, it is beautiful.

This morning I am finishing a Western blot I started yesterday. I wish I could say the data will be earth shattering but it is just an attempt to find a good loading volume. My protein assay isn't working reliably so I am doing it empirically. When this is done, I am going to teach a person in the surgery department how to perform a rat surgery - kind of ironic, isn't it? It is actually a very basic procedure. We're just making a pouch under the skin between to shoulder blades, placing an estrogen pellet in the pouch and suturing the wound back up. The only tricky part is getting the anesthesia right. I really enjoy doing animal surgeries. They are challenging and satisfying when you get them right. Also, the amount of data that you can get is well worth the work. The only problem I am experiencing is that I am becoming allergic to either the rats or their bedding.

You have got to be kidding me.

After receiving my MS in Biology at Idaho State University, I came to Iowa to complete a PhD. After two years in a great program, I left. Partly because I just didn't have the time to devote to my studies. As the father of 6 children, I felt I needed to spend more time focusing on my young family rather than ignoring them in order to further my own desires. I probably could have stayed in and finished with a mediocre thesis but I have too much respect for the degree to have done that. I write this because I have seen so many people come out of graduate programs with doctoral degrees that really didn't deserve them. Case in point. We have a post-doctorate fellow in our lab who has been with us now for 7 months. I still have to remind her to write in her lab notebook. My boss asked me to teach her how to column purify protein. I first had her transfect a flask of HEK 293 cells (a well established kidney cell line) with a DNA construct containing our gene of interest. The cells would then use their machinary to produce our protein. The protein was engineered to have what is called a histidine tag expressed with our protein. This enables us to purify our protein by running the cell lysate over sepharose beads that have Cobalt attached to them. Cobalt and histidine form a chemical bond which allows you to keep your protein of interst bound to the beads while everything else just passes on by. After washing the beads a number times to remove anything that is bound in a non-specific manner, you can then elute your protein of interest by washing the beads in a solution containing Imidizole and if the Voodoo king is smiling on you, you will have purified protein. Anyway, this is a fairly long and drawn out proceedure. After guiding our Post-doc through the proceedure, having her run the samples on a polyacrylimide gel, and then staining the gel with silver to see the protein, I was pleased to see the elution profile I had predicted for her. I then told her that since the silver stain was non-specific, I wanted her to run the gel again, transfer the proteins to a nitrocellulose membrane and probe the membrane with an antibody specific for our protein. I told her that if we saw the same profile, we could be assured that we had pulled off the purification. She looked at me blankly and said, "How can we do that? There is no his-tagged protein in that solution." I stared at her for a moment, not really sure what she meant, but assured her that this is what we needed to do. She again told me that wouldn't work. I was still scratching my head. She informed me that she had done the entire experiment with a solution from a non-transfected flask of cells. She tried to tell me that this was a good negative control. I continued to stare at her not believing what I was hearing. Why would you even try to purify a protein that you knew wasn't there in the first place? That is not a good negative control - especially since we got a result.

I find it hard to believe that someone with a PhD in the biomedical sciences never once thought to question me or the procedure. The difference between a PhD and a lab tech is that a PhD is supposed to be able to think critically. This Post-doc has yet to rise above the level of a newly minted lab tech. I am sure she will eventually but it is getting tiresome waiting.

Hey

First to explain the title of my blog. I work in the Otolaryngolgy department (the cochlea part) at the University of Iowa (the corn part) where I study the cell physiology of hearing and hearing nerve-related tumors. Most Americans seem to confuse Iowa with Idaho or Ohio. Why? I am not sure. Maybe they are stupid or too self absorbed to realize that there are states in between New York and California. Those who know something about Iowa think of corn (which comes in ears) and hogs. And rightly so. Iowa is a huge agricultural state. What people don't realize is that Iowa is also home to one of the best public university's in the nation - at least from a biomedical standpoint. My department is actually ranked the second best in the nation behind Johns Hopkins University. In additon to having top notch clinical care, we also have great research efforts going on. In my job as a lab manager, I have what in my mind is the best of all worlds. I get to be involved in the day to day administration of the lab (which does have its drawbacks) as well as being involved in training new lab members and doing my own research at the bench. I am going to use this blog as a way to catalog some of the stuff that goes on around here - both the good and the frustrating. Anybody in research will tell you that being frustrated is one of the side effects of the job. Having said that, I really do love my job. I have an incredible boss and work with some very bright people.