PREFACE iii

Preface

The following manuscript is the product of my efforts to understand a fraction of the

mind of God.

From the beginning, I had the great fortune of adopting one of my predecessor’s final

projects as my first project. Working on this project became a milestone in my studies,

not only as an initial project, but more so because of the experience of working with an

adept NMR scientist. The preliminary hypothesis of this study was to observe changes in

murine tumor oxygenation with radiotherapy. In preceding works, it was shown that the

spin-lattice relaxation rate of perfluorocarbons was directly proportional to the dissolved

oxygen tension. This relationship between fluorine relaxation and oxygenation, along

with NMR imaging techniques, allowed for evaluation of therapeutic interventions using

perfluorocarbons.

During this time, a parallel experiment was in progress to observe the effects of

oxygenation on the diffusion of water molecules in murine tumors. Other groups had

shown that there was a highly positive correlation between tumor oxygenation and the

apparent diffusion coefficient (ADC) of water in the tumor. These findings were

troubling because intuitively, if there were any correlation between tumor oxygenation

and ADC, we would expect to see a negative rather than a positive correlation. To
PREFACE iv

resolve this issue, we undertook the project of experimentally correlating tumor

19
oxygenation using F imaging and 1H ADC mapping techniques. This proved to be a

rewarding experiment because our intuition served us correctly. Despite the findings by

other groups, our studies showed that no apparent correlation existed between ADC

values and tumor oxygenation.

Studies in animals brought up questions about the behavior of water in the brain

following the onset of stroke. During cerebral ischemia, ionic imbalances in the brain

result in changes in cellular water distribution and give rise to edema (cellular swelling).

Although it is clear that the properties of diffusing water are correlated with the swelling

of cells, it was difficult to understand the intricacies behind the relationship of water to its

surroundings. The answer to this basic question of what is happening to the water in a

complicated two-compartment system has been one of the motivating factors for my

study of water in a model system of yeast cells. Studies of two-compartment systems by

others have shown non-monoexponential diffusion signal attenuation, which has been

assumed to be because of the different compartmental contributions. Although this may

be a possibility, previous works have shown that even in single compartment systems,

non-monoexponential behavior could be observed simply as a result of the tortuous

diffusion pathways of water in the interstitial space. This observation in single

compartment systems cast doubt on the interpretation of the data from the two-

compartment system. Not only are there compartmental effects on diffusion, but the

diffusion behavior is also confounded by restriction effects. One of the premises behind
PREFACE v

the work done in yeast-cell suspensions was that a deconvolution of the compartmental

effects could provide valuable information on the behavior of water. With this goal in

mind, a majority of my research efforts was devoted to investigating model systems for

understanding the diffusion behavior of water in a two-compartment system. Fortunately,

a well-characterized system was available in yeast-cell suspensions. The yeast

experiments should serve merely as a stepping stone to other experiments which further

characterize the model system and, ultimately, carry over to in vivo experiments to

understand the behavior of water during ischemic brain injury.

The last experiment in this dissertation deals with diffusion of water in excised rabbit

Achilles tendon. The initial set of experiments on tendon gave very promising results and

eventually led to a Master’s thesis for one of my colleagues. Unfortunately, further

investigation into the behavior of the tendon revealed that these observed effects were no

more than an artifact of the storage medium. The initial discovery of this fact was

disappointing, but at the same time opened up another avenue for the direction of these

experiments. In wrestling with the logistics of measuring NMR properties of tendons,

most of the groundwork has been laid for future studies. Although these results are not

completely revealing, we are still of the belief that the viscoelastic properties of tendons

change with tensile load. Whether this phenomenon is observable via NMR is still to be

determined.
PREFACE vi

Although diverse in nature, all of these experiments have taught one common lesson.

Ask the good question and seek the right answer. I hope that the writings herein will be a

reflection of the work done to seek the right answer.

May God bless the reading of this dissertation.