Experience is the best teacher. "The great Roman leader Julius Caesar recorded the earliest known version of this proverb, 'Experience is the teacher of all things,' in 'De Bello Civili' (c. 52 B.C.). Reference.
This experience stems from the idea of lipid peroxyl radical production of fatty acids by lipoxygenase using an electron spin resonance spectrometer (ESR). While we had other experiments lined up while visiting the University of Iowa Medical Center, this particular one was our high risk one given the limited background preperation in understanding the chemistry of lipoxygenase. Some work (quite admirable) has been iniated by Ron Mason from the Laboratory of Molecular Biophysics at the National Institute of enviromental health Sceinces in the Research Triangle Park of eastern North Carolina. Understanding these chemical species is important as it is proposed they are intermediates in biological functions proposed to be mediators of hydroperoxide dependent oxidations related to human diseases. There have been several mechanistic models that perdict this [de Groot, Garssen, Vliegenthart and Bolding (1973) Biochem. Biophys. Acta 326, 279-284; (1975) Biochem. Biophy. Acta 377, 71-79; ... to more resently Gardner, H. W. (1989) Free Rad. Biol. Med. 7, 65-86; reference Mason, R. (1989) J. of Bio. Chem. 264, 20968 - 20973]. Now that I've set the stage for the enduring nature (and understanding) of our interests it is important to note true learning is derived from experience and hence this blog entry.
Four researchers (with varing skill sets) entered the laboratories of the University of Iowa armed with linolenic acid (a 2-bis structured creature - one of the two hydrogens at the bis junction is thought to pop off leaving behind it's electron to under go radical chemistry) disolved in absolute alcohol and then diluted in nanopure water, and some lipoxygenase attached to some agurose jel beads. Our team is prepared .. so we think. Once we loaded the ESR with our flat cell filled with lipoxygenase agerose beads a syringe pump was used to pump the linolenic acid solution. We wait. Nothing. Instrument adjustments were made ... still nothing detectable. Welp. Yeah.
The two undergraduate researchers were devastated without tangible results for which we would have the predicted signal. If for no other reason this gives rise to further refine our understanding of the previous research and trouble shooting our application of the first round in the scientific method. It is within this cyclical function we now find ourselves. Something as simple as some iron containing molecule contamination has probably reduced (degraded) our linolenic acid (this is my thought) or there is another source we haven't considered which has already reacted the linolenic acid ... or for that matter it is plausible the lipoxygenase never bonded to the agerose beads. eh ... who knows. The important thing is that this lesson in perserverance provides us with potential insight to the pure chemistry we are interested in revealing. Armed with some experience, we are ready to try, try again. Science worth doing isn't typically easy.
Many thanks to Dr. Brad Sturgeon of Monmouth University, Illinois, and to Dr. Garry Buettner of the University of Iowa, Iowa.