Professor Sparks was excited.

“Good, we finally have a clear difference of opinion here.” He continued quickly. “Joe, you did not think that the conclusions were supported by the data. What data would you personally want to see to support those conclusions?”

I was on the spot and all alone on my opinion, but felt I needed to defend it.

“I think they need to do some experiments on live cells or tissue. All of their experiments were done with fixed cells, which are dead. If the conclusion was that a motion occurred that was like a muscle contraction, they did not show contraction. All they showed was differences in the cell shape before and after. They did a lot of manipulations after the cells were dead and during the fixing, so I feel that the results could be an artifact of their methods and not physiology of the cells.”

Nodding, he opened the question up to the class, “Who felt that the methods were solidly supporting the conclusions and why?”

He pointed to Ken and asked for his opinion. Ken pointed out that the differences seen between the groups of cells in the experiments were large ones and that for the differences to be so big the conclusion would have to be that the cells controlled blood flow. Several of my classmates agreed with Ken and eventually with further discussion were all nodding their assent, that it was a good paper.

Professor Sparks let that sink in and then reconciled the discussion for the class. “This paper is considered a seminal paper in physiology,” he said, meaning that my criticisms were wrong. “However,” he went on, “it is seminal because many scientists pointed out some of Joe’s criticisms as well as others and those experiments in living cells were eventually carried out. So both Ken and Joe are correct. It is a solid piece of physiology but it needed more work to verify the conclusions. Good job, everyone.” I think all of us in the class realized that a good opinion that has reasonable justification is a valid opinion. At the cutting edges of science there may not always be a right or wrong, but a difference of opinion and/or a consensus of opinions. Maybe we had been missing the point when my classmates and I agreed on one perspective and marched like lemmings into the class. After that day we no longer compared notes on our reading assignments and let differences of opinion be a catalyst for further discussion. It actually was a lot more fun if people disagreed and we could engage in friendly debate on the subject. As a group we were teaching each other to think as individuals and to respect differing opinions.

A lot of thought went into my Ph.D. project. I wanted to study vascular smooth muscle because there were so many diseases of the arteries that I could do research on. I wanted to learn about its function; contraction and relaxation, but I also its chemistry or the inner workings of the muscle. Because I had worked a lot on the nuclear magnetic resonance machine for my master’s, I decided that I would use that to study the chemical kinetics of at least one enzyme in the artery muscle. I would study pig arteries because pig arteries were very much like human arteries. I did not want to have to get human arteries at the hospital. Michigan State had a slaughterhouse on campus and the tissue from pigs killed for food was also available for research. There were also other slaughterhouses near campus that could be a source of research material as well.

The graduate students took classes together and in these classes we learned the theory of enzyme kinetics. There is a lot of math and many formulas to memorize in enzyme kinetics. The math was used to predict and explain how and why two chemicals interacted during a chemical reaction. We often learned how the equations were determined by scientists doing research, but all too often we learned very little as to how to use them. Sometimes we learned equations and wanted to use them. A colleague in my class named Tim Denerall fell in love with an equation. He described it as ”beautiful” and extolled its virtues for doing enzyme kinetics. The equation to describe the velocity and inhibition of reactions uses the formula[1]:

              Vmaxa [A]    + Vmaxb   [B]

Vt =                  Kma                         Kmb. 

              1   + [A]   +   [B]   

                      Kma       Kmb

The beauty of this equation, he said, was in its simplicity and utility, although that might not be evident to people who do not do enzyme kinetics, I have to admit I have personally never fallen in love with an equation, and sometimes I have to work hard to remember them because I tend to dislike them, but they can be useful for people doing biochemical studies in graduate school.

For my research I needed something that could be studied in vascular smooth muscle that would be able to fuse all my interests. I needed a subject that was important to chemistry and the physiology of arteries that would be testable in the nuclear magnetic instrument and that could eventually be of great help to people. After much thought and reading of the literature on the subject, I decided to study creatine and creatine kinase. Creatine is a molecule important to all muscles and the brain. It is important to athletes as a dietary supplement and the main function of creatine in the human body is to be acted on by creatine kinase, whereupon it makes and stores energy that is central to all muscle and brain work. I would study the arteries and determine how creatine and creatine kinase were used chemically in the muscle to aid muscle function.

I outlined my idea for the project to Dr. Dillon and he said yes. He actually said it sounded like a good idea. I was relieved and disappointed at the same time. I had read tons of books and papers on the subject and was prepared to justify my thinking with backup material in response to any questions or arguments he might have. But none of that was needed and I was now on the Ph.D. treadmill.

[1] Taken from Page 113 of Enzyme Kinetics, Irwin H. Segel, © 1975; John Wiley and Sons Canada.