Water Structure Changes in Dilute Solutions of Physiologically Active Substances

Kenneth S. Marsh, Ki Brissey, Su-il Park

Abstract

100 samples of dilute solutions of physiologically active substances were prepared in water and a vicinal water solution that simulated cellular interfaces. The solutions were then scanned in a medical MRI. The results demonstrate that dilute solutions of these substances yield a wide range of response suggesting that the water protons are experiencing variations in the water environment. The vicinal solutions exhibited more contrast than the corresponding bulk water solutions. The suggestion is that physiologically active substances modify water structure, especially interfacial water, and that this influence on water structure is part of their physiological effect.

Introduction

This experiment is designed to investigate the hypothesis that pharmacological action is accomplished, at least in part, through water structure changes. A necessary (but not sufficient) manifestation of this hypothesis would be observance of experimental changes in measures of water structure through the influence of low concentrations of pharmacologically active substances.

It is known that the organization of molecules in the water matrix changes in space and time and can be modified through interactions with macromolecules, ions, and other chemical compounds. The exact nature of these changes is not known, and includes numerous forces including hydrogen bonding, van der Waals forces and dipole effects. For lack of a better term, we will employ “electronic” interactions to imply influences of the electron cloud. The existence of the different environments is exhibited in Magnetic Resonance (MR) spectra. T1 relaxation times vary with the special and electronic environment surrounding water protons, and are therefore used to present differences in that environment, both in chart form (nuclear magnetic resonance spectra) and graphic form (MR imaging or MRI). The sum total of all interactions on the water protons effects its ability to realign with the magnetic field after the radio frequency pulse is removed, which presents subtle changes in the water environment with optical density differences in the MRI. We will refer to these different environments collectively as water structure with no implication that we know the configuration of these structures. We will, however, demonstrate that water structure is influenced by very low concentrations of physiologically active substances.

 

Background

Michael Tracey (1964) determined that pharmacologically active substances which are known to be stimulants acted upon the rheology of wheat dough systems as if additional water had been added to the system during dough development. Depressants acted on the dough as if water had been removed during dough development. These actions were obtained with drugs at physiological levels, i.e. levels below which colligative effects would be anticipated. Tracey defined stimulants as water structure breakers and depressants as water structure makers.

Although not specifically stated in Tracey’s study, the wheat gluten experiment represented a system characterized by interfaces. The interfacial effects of water have been well studied by Walter Drost-Hansen who defined a series of anomalous interactions which occur at interfaces. The term “vicinal water” describes water in proximity to a surface, which was found by Drost-Hansen to be any substance with a molecular weight exceeding 3000-5000 Daltons. The occurrence of these anomalous behaviors in the vicinity of a surface, regardless of the surface composition, was described by Drost-Hansen as the paradoxical effect. He went further to suggest that most, if not all, of the water in cells would be sufficiently close to a sufficiently large molecular weight entity to act vicinally.

Plant systems cannot promote survival by a fight or flight response typical of the animal kingdom. As a result, plants must combat environmental and other attacks chemically. Plant systems have evolved a cornucopia of chemical agents which have beneficially impacted their survival. Humans have extracted many plant alkaloids and found them to be physiologically active in animals (including humans) and are the basis for many drugs.

If water structural changes are vital to life processes (as described by Drost-Hansen) and modifications of water structure are implicated in drug action (as suggested by Tracey), then it follows that drugs should exhibit modifications of water structure. This paper is designed to study this premise.

Our intent is to study MR images of very dilute drug concentrations in both standard and vicinal solutions.

 

Experimental Design

The concept under test is that physiologically active substances in low concentration will demonstrate a change in water structure as measured through MRI. Since the biological systems exist in a vicinal environment, vicinal and bulk water environments will be employed.

 Sample Choice

Physiologically active substances are compounds which demonstrate effects on life processes at low concentrations. Stimulants, depressants, humectants, chemotherapeutic agents, vitamin C, hormones, and organic acids were chosen for study. Stimulants and depressants were chosen to expand on the work of M. Tracy. Vitamin C was included because of previous work by the WIS2H Institute (unpublished) which found MRI scan of vitamin C to reveal an unexpected change in water structure as indicated by a reduced T1. Additional organic acids were included to determine if the dramatic response of ascorbic acid is an effect of unusual properties of ascorbic acid itself, or simply a response to characteristics of an organic acid.

Humectants are substances which modify water holding capacity. Humectants were included to obtain an alternate measure of humectant activity through MRI and also to test substances known to have a direct effect on water. The data will also provide a forum for further studies which relate water structure and humectant activity to biological reactions and growth. Since water control has been a traditional means to preserve food products by limiting microbial growth, microbial growth studies are planned.

Chemotherapeutics were included to explore the hypothesis from WIS2H that changes in water structure must precede malignant (or any) growth. A random effect on optical density of the MRI would suggest that water structure influence is not a factor in chemotherapeutic response; any non-random behavior would provide evidence to support the hypothesis, with degree of non-random behavior a factor. Water structural changes are necessary, but not sufficient for proving the hypothesis. Therefore this work is exploratory.

Cortisone and progesterone were included because hormones have greater influence (in terms of concentration effects) than most drugs, and also influence water balance in the body at times of stress.

 

Solution Preparation

Much literature suggests that interfacial water behaves differently from bulk water (Drost-Hansen & Clegg, 1979; Franks, 1972 – 1979; Pollack, 2001). Therefore, for the purposes of this study, both water and interfacial water systems were chosen. The interfacial system was modeled by employing a polymer system with a molecular weight greater than 10K Daltons, which was suggested by W. Drost-Hansen () as above the limit for “vicinal” or interfacial differences. Polyvinylalcohol (PVOh) was chosen because it is water soluble, contains a relatively simple and non-reactive chemical structure, and possesses a sufficiently large molecular weight. The molecular weight of synthetic polymers, unlike proteins, is not a specific value but a range. DuPont Elvanol, Grade 90-50 was chosen for this study. Elvano 90-50 was found to posses a molecular weight of 29,700 Daltons by the numerical method (which emphasizes the lower MW components) and 55,800 Daltons by the weight method (which emphasizes the higher MW components), which is considerably larger than the minimum required for vicinal effects. The concentration of PVOh was calculated to allow 50 water molecules for each repeating unit of the polymer matrix, specifically -(CH2-CHOH)-. This structure possesses a molecular weight of 44, so 44 grams of PVOh were used for each 18 MW * 50 water layers or 900 grams of water. The water in this polymer solution, therefore, will all be statistically well within 50 molecular distances of a polymer chain.

 

Physiological Dosage

Each drug was prepared in standard dosage forms and equi-molar concentrations into both water and PVOh solutions. The “standard” dosages were obtained through dosage recommendations in product literature or through the Physicians Desk Reference (PDR). These dosages are typically the available pill sizes for the drugs. The dosages were converted into concentrations through the following procedure. Dosages are typically calculated in terms of mg/kg body weight. For convenience, manufacturers prepare standard dosages by calculating the dosage required for an average person, typically a 150 pound male. An average built 150 pound male has a blood volume of 4.77 liters (Luisada, 1959). With a simplifying, working assumption in which the active ingredient is uniformly dispersed in the bloodstream (and not in tissue), we applied the pill dosage into 4.77 liters for our “physiological” concentrations. It is recognized that some drugs are metabolized into an active form. Such mechanisms are not incorporated into this study.

The next step was to choose a mean concentration, with the absolute realization that no such meaningful average drug dosages exists. Different drugs act at different concentrations, and any particular value would be a gross over-simplification of drug action. Furthermore, the above concentrations are based upon dosage, not molecular weight. However, the purpose of this study is to compare action on water structure systems, as defined by MRI, and equal concentrations are desired for direct comparison. Molar concentrations were calculated for each test substance and a median value was chosen for the experimental protocol and rounded to concentration of 0.0005M.

The resulting protocol consisted of 19 physiologically active compounds, each in standard dose and a concentration of 0.0005 molar, and each in water and PVOh solution. Some compounds were tested with various concentrations to represent different dosage levels. Pure water and PVOh solutions were incorporated into the test. A total of 100 samples plus four index controls (distilled water) were scanned.

Many of the compounds were available in pure form (sugars, caffeine, organic acids, glycerine). Analgesics and pharmaceuticals were requested in pure form. Those which could not be obtained pure (Vioxx, Phenobarbital, some chemotherapeutic agents) were used in pill form. The pill was ground in a morter and pestle, and a proportionate quantity was weighted with the same ratio of dosage to pill weight in order to compensate for the weight of the binders.

 

Experimental Procedure

A stock “vicinal” solutions was prepared by dissolving 44 grams of Elvanol 90-50 into 900 ml of distilled water. Distilled water from the same source was use for the “bulk” water samples. Appropriate weights of each active substance were added into both distilled water and vicinal water to prepare the solutions of the correct concentrations. Each test solution was placed into a 50 ml graduated conical polypropylene tube with cap (Becton Dickinson Bluemax 2098) in an expanded polystyrene rack. Each rack held 25 tubes. To assure that the films were not inadvertently reversed during viewing, a 1/2″ diameter polypropylene test tube was filled with distilled water and placed in one corner of the rack. This tube provided a second distilled water standard and served as an index for reading the films.

The racks were placed on an expanded LDPE base (Dow Ethafoam 220), which was cut to a rectangular block of 10.5 by 6.125 by 3 inches high. The bottom long edges were optimally radiused to 1.25 inches to allow the base and rack to correctly position the sample tubes across the horizontal diameter of the Atlas Head/Neck Vascular Phased Array Coil. However, truncating the edge with a straight 45° cut, 1.25 inches horizontally and vertically from the bottom edge was proven adequate.   The entire rack system was scanned in a Polaris 1.0T MRI Magnetic Imaging system through the cooperation of the Department of Radiology at Oconee Memorial Hospital. Four sets of 25 samples were tested.

The first set was scanned for T1 weighed, T2 weighed, Proton Density weighted, and dual T2 /Proton Density weighted responses. The greatest contrast differences between samples was observed with the T1 weighted protocol so T1 was chosen for the study. The differences in T1 were reflected as differences in optical density of the individual images.

The resulting cross-sectional circles from each tube on the film (coronal slice, representing each sample) were evaluated for optical density. Two densitometers were tried, but it was found that the fine structure in the images yielded unacceptable variance in the densitometer readings. An integration over the image was required, but not available through the optical system of either densitometer. Visual inspections were therefore performed in which relative optical densities were evaluated before the identity of the samples was known to the evaluator. Comparative densities were recorded on a chart which included circles representing each sample, and a place to record the image and plate number from the MRI film.

A quantitative evaluation of optical densities was obtained through a colorimeter (Minolta Chroma Meter CR-3000), which provided a larger sample area, and therefore integrated the density for each sample.   The “brightness” was obtained through the L-value of the CIE L* Color Space.

The MRI scans of the samples exhibited a non-uniform optical density even though the samples were homogeneous. In order to investigate if the cause was a dielectric effect of polypropylene, four samples of PVOh were incorporated into the fourth experimental set in glass rather than polypropylene holders.

 

Results

The results of the MRI scans are presented in Figures 1 – 4, with the densitometer values for each sample presented in Tables 1 – 4. Controls of pure distilled water and PVOh solution are included.   Table 5 presents the four scans in a single table, ranked by increasing image density. The 100 samples scanned represented concentrations of physiological substances from 5.8 x 10-6 to 2.3 x 10-3 Molar and exhibited optical densities ranging from 66.47 to 26.75, with higher densities representing a darker image.

 

Discussion

The optical density, both visually (Figures 1 – 4) and numerically (Tables 1 – 4) illustrate that relaxation times of water protons are influenced by low concentrations of pharmacologically active substances. Observations must be taken as tendencies rather than absolute comparisons for a number of reasons. First the field density in the MRI is not uniform. The center of the coil shows brighter images than the periphery. With medical images, the contrast relationships provide valuable information for diagnoses, but the optical densities vary across the film for similar water environments. Therefore, our quantification of optical densities must be evaluated in this context. Direct T1 measurements could have been obtained on a NMR spectrophotometer, which was not available for this study. Proper shimming of the magnet would also reduce the varience.

Set 1                            DI-Table 1                              DI-Photo Set 1

The T1 weighted images exhibited larger contrast between the samples than T2 weighted or proton weighted images. With T1 weighting, the brightest images appeared to be vitamin C in PVOh for the 250, 500 and 1000 concentrations. The vitamin C 1000 sample appeared light, but darker than the other vitamin C in PVOh. The next noticeable group was tartaric acid in PVOh (both 500 and 1000) and caffeine in PVOh (both 50 and 200). The next darker group is citric acid 60, 250, 500 in PVOh, with the citric acid 1000 again slightly darker. Next is vitamin C in water , then citric acid in water, caffeine 200 in water, tartaric 500 and 1000 in water, and final solution is caffeine 50 in water. The pure PVOh sample appears approximately equivalent to vitamin C 500 in water. Water in pure form was darker, but more so with the sample in the smaller tube near the extent of the field. Pure water, as expected, exhibited a longer relaxation time, suggesting that samples are essentially contributing some structure in the water environment..

Ascorbic acid seemed to have a stronger effect on water structure than tartaric and citric acids. This is unlikely to be strictly a manifestation of properties of an organic acid because the Pka values are comparable. Molecular weights are comparable (ascorbic acid, citric acid and tartaric acid have molecular weights of 176.12, 192.12 and 150.09 respectively) but the configuration of the ascorbic acid appears capable of a stronger influence on the surrounding water. This supports the hypothesis of a relationship between water structure and physiological activity.

Our studies exhibited action of caffeine which agrees with Tracey’s work, i.e. demonstrated evidence of a water structure maker. Although the Phenobarbital was considerably less active in the MRI, it still reduced the optical density in the MRI compared with pure water. This finding does not appear to agree with Tracey’s observation on first look. Three factors complicate the observation. First, the caffeine was available in pure form but the phenobarbital was only available in pill form, with the standard binders and sugars.   These additional components could alter the observations. Secondly, water samples in different locations exhibited different responses because of field effects in the MRI. One water sample was more active than the Phenobarbital; one was less active. Thirdly, the observed differences should be related to vicinal water. Although Tracey did not state this directly, most or all of the water in a wheat gluten dough systems would be expected to be vicinal. Therefore, the standard for comparison should be with vicinal water, not bulk water.) Hazlewood et al. (1974) reported that at least three fractions of water exist in cells, all of which exhibit faster relaxation times than bulk water. Another interpretation, therefore, is that wtare-structure breaking and making is relative to a value other than that of bulk water – further support for the vicinal water explanation.

Set 2                            DI-Table 2                              DI-Photo Set 2

Set 2 was designed to address the effects of analgesics which also serve as anti-inflammatories. Aspirin is used as a blood thinner and is therefore employed because of known effects on aqueous solution properties. Furthermore, aspirin may also serve as a water ‘thinner’ in the plant kingdom. Mansfield and Marsh (1995) measured voltage potential between willow bark and a reference stake approximately three meters from the base of the trunk. The voltage potential increased during the autumn months in direct correlation with acetylsalicylic acid production, suggesting that the aspirin production was facilitating flow of water as the temperature dropped in preparation for the dormant winter. Theses effects are complex, but this effect is consistent with structure modifications and could be a contributing factor for water transport in trees.

Other anti-inflammatory also exhibited influences on the water structure as shown by density variations in the MR images.

Set 3                            DI-Table 3                              DI-Photo Set 3

Set 3 evaluated effects of humectants to explore water structure differences projected by the WIS2H Institute based upon the work of J. E. L. Corry. Corry (1975) performed growth studies with Salmonella Typhimurium in which water activity was controlled with a variety of humectants. The results showed that different humectants provided different growth suppression at the same water activity. The generally accepted principle is that microbial growth is a function of water activity, and therefore water activity is used as a standard for food safety. This growth study suggests that water activity (the partial pressure of water above a solution divided by the saturated vapor pressure) is not the primary effect. If water structure were implicated, then humectants in dilute vicinal solution might exhibit action in the same rank order as the Corry study. Sucrose exhibited the most dramatic reduction in T1 relaxation, followed by dextrose followed by glycerine, which follows the growth suppression found by Corry. The implication is that the effects on these humectants on water structure relates to the availability of the water to support microbial growth. Since humectants are employed for food preservation, the effects of humectant activity on both water structure and microbial growth deserve further study.

Set 4                            DI-Table 4                              DI-Photo Set 4

Set 4 explored the effects of chemotherapeutic agents and also included two hormones. These agents were only available in the pill form, so the solutions contained binders. Vioxx (from pill form), and fructose (pure) were included in this sample set. The set also included PVOh solutions in glass tubes to determine if the patterns observed in previous samples resulted any interactions with the polypropylene sample tubes.

The chemotherapeutics and hormones act at lower dosages than most of the previously mentioned compounds. The hormones progesterone and cortisone were tested at 5.8 x 10-6 and 8.7 x 10-6 Molar concentrations respectively as well as the “standard” concentration of 5 x 10-4 Molar. These low concentrations yielded responses that still exhibited differences from pure water.

Combined Tables 1-4

 

Discussion

The observation of dramatic differences in MR response in low concentrations (between 10-3 and 10-6 Molar) suggests that these pharmacologically active substances are exerting an influence on water protons at concentrations far below levels that would be expected for colligative effects. This supports the hypothesis that physiological activity may be related, at least in part, to effects on water structure.

It is significant that water solutions of pharmacologically active substances exhibited more subtle differences than similar concentrations of these active substances in vicinal solutions. This evidence suggests that active substances preferentially influence structural alterations in vicinal solutions, and therefore modify the interfacial relationships between water and solutes. The decrease in the optical density of the MR scans compared with bulk water solutions suggests a more structured water environment in which energy from the radio frequency pulse in the MR is more rapidly dissipated through electronic interaction between the water protons and the matrix. This is consistent with the accepted theory of NMR response in which so-called “bound” structures provide a path to dissipate energy and thereby yield a shorter relaxation time for the molecule in question to realign with the magnetic field after the radio frequency pulse ceases. We make no claim to know the morphology of the structure, but it is entirely reasonable that active substance create structure. It deserves mention that the term “bound” is a misnomer because radio labeled water was found to move, but the term is used to describe a tendency for water accumulation as opposed to a bonded structure. Hazlewood’s findings of three fractions of cell water with faster relaxation times also supports the idea of active structuring.

The fact that the effects are more subtle in bulk water may explain why water structure action of pharmaceuticals is not a typically evaluated function. Considering that “most if not all of the cellular water is vicinally hydrated” (Drost-Hansen, 2006), physiological structuring is significant in vicinal water, It is therefore reasonable that the physiological activity is related, at least in part, to the effects these compound exert on water structure. If verified, these influences could provide valuable insights into drug action which could promote development of improved agents.

A somewhat simplistic classification of physiological activity can be described as follows: food offers physiological activity in high concentration, drugs offer activity in low concentration, and hormones result in physiological activity with very low concentrations. The hypothesis that physiological activity is related to water structure is supported by the findings that compounds with known activity demonstrated alterations in water structure as imaged through MRI. Furthermore, action was observed with low concentrations of drugs and very low concentrations of hormones.

The field in the MRI varies across the sample area, as illustrated by multiple samples placed in different locations within the test chamber of the MRI. However, the variation sample to sample were considerably larger than comparisons with similar samples. The findings therefore support the hypothesis that the physiologically active substances also modify water structure. These finds are necessary for the hypothesis, but not sufficient to prove the hypothesis. This study is therefore offered to open exploration into the relationships between physiological activity and water structure with the hopes that findings will improve our ability to identify more effective agents.

Acknowledgements

The authors wish to thank the support of the Woodstock Institute for Science in Service to Humanity for sponsoring this study, Occonee Memorial Hospital for their support and access to the MRI, and Hubbard Young Pharmacy for valuable assistance with the pharmaceutical choice and properties. We also want to acknowledge the support of Du Pont for the Elvanol, and McNeil Consumer Healthcare, Pharmacia, who supplied their respective pharmaceuticals, and Research Biochemicals International for supplying Fluoxetine in pure form so that we could conduct this study without influences of binders and other agents which exist in the tablet form. Finally, I wish to thank Brookfield Instruments Company for use of a viscometer which could measure low viscosity solutions used in this study.

 

References

Corry, J. E. L., 1974, “Effects of Sugars and Polyols on the Heat Resistance of S. Typhimurium”, J. Applied Bacteriology, 37, 31.

Drost-Hansen, W. has published 50 years of research on Vicinal water. One summary is “The Role of Vicinal Water in Cells”, accepted May 3, 1994 for publication in Medical Hypotheses, D. F. Horrobin, ed., Churchill Livingston Medical Publishers, Edinburgh, U.K.

Drost-Hansen, W., 2006, “Chapter 9 – Vicinal Hydration of Biopolymers: Cell Biological Consequences”, in Water and the Cell, edited by G. H. Pollack, I. L. Cameron and D. N. Wheatly, Springer, Dordrecht, The Netherlands.

Drost-Hansen, W. and Clegg, J., 1979, Cell Associated Water, Academic Press,

Franks, F., 1972 – 1979, Water – A Comprehensive Treatise on Water, (Volumes 1 – 7), Plenum Press

Hazlewood, C.F., Cleveland, G., and Medina, D.  “Relationship Between Hydration and Proton Nuclear Magnetic Resonance Relaxation Times in Tissues of Tumor-Bearing Mice:  Implications for Cancer Detection”.  J. Nat. Cancer Inst. 1974, 52:1849-1852.

Luisada, A.A., 1959, in Handbook of Circulation, Dittmer and Grebe, (Eds.), Saunders, Philadelphia, page 112.

Mansfield, S., Marsh, K. 1995, Personal Communications.

Physician’s Desk Reference

Pollack, G. H., 2001, Cells, Gels and the Engines of Life, Ebner & Sons, Seattle, WA.

 

Table 1: MRI Drug study – Scan set 1       

Scale: Structured =light = higher L value; Free = low L value

Compound                Solvent            Dosage           Molarity         L Value       Position

Tartaric acid               PVOh              1000              1.4 x 10-3         66.46                     9

Caffeine                       PVOh                200              2.2 x 10-4         66.37                    21

Vitamin C                    PVOh                250               3.0 x 10-4       66.22                     11

Vitamin C                    PVOh                  60               7.1 x 10-5         66.11                      4

Vitamin C                    PVOh                500               5.9 x 10-4         66.06                    17

Vitamin C                    PVOh              1000               1.2 x 10-3         65.04                    23

Tartaric acid                PVOh              500                7.0 x 10-4         64.97                      2

Caffeine                       PVOh                 50                5.4 x 10-5         64.01                    15

Citric acid                    PVOh                 500               5.5 x 10-4         55.23                    19

Citric acid                    PVOh                 60                6.5 x 10-5         53.39                      6

Vitamin C                    water            1000                1.2 x 10-3         52.36                    22

Citric acid                    PVOh              1000               1.1 x 10-3         49.86                    25

Citric acid                    water                  500               5.5 x 10-4         49.51                    18

Vitamin C                    water                  500               5.9 x 10-4         49.08                    16

Vitamin C                    water                  60                7.1 x 10-5         48.03                      3

Vitamin C                    water               250                3.0 x 10-4         47.83                    10

Citric acid                    water               1000                1.1 x 10-3         46.63                    24

Citric acid                    water                  60                 6.5 x 10-5         45.82                      5

Citric acid                    PVOh              250                 2.7 x 10-4         45.51                    13

Pure water                                                                     55                     43.45                    12

Caffeine                       water               200                 2.2 x 10-4         40.70                    20

Pure PVOh                                                                     48.9                  37.87                      7

Tartaric acid                water               1000                1.4 x 10-3         36.86                      8

Caffeine                       water               50                  5.4 x 10-5              33.32                    14

Tartaric acid                water               500                7.0 x 10-4            33.13                      1

Water index                                                                      55                   29.49               Index

 

Background = 25.52            Clear Background = 66.06

 

Table 2: MRI Drug study – Scan set 2                   

Scale: Structured =light = higher L value; Free = low L value

Compound                  Solvent            Dosage            Molarity         L Value            Position

Citric acid                    PVOh              .0005M           5.0 x 10-4         65.02                    10

Acetomenophen          PVOh                   650             9.0 x 10-4         64.43                    11

Acetomenophen          PVOh                 1000             1.4 x 10-3         64.13                    17

Caffeine                       PVOh              .0005M           5.0 x 10-4         63.92                    16

Fluoxetine                   PVOh                   60              3.6 x 10-5         62.89                    21

Acetomenophen          PVOh              .0005M           5.0 x 10-4         62.39                    22

Ibuprophen                 PVOh              .0005M           5.0 x 10-4         62.21                    23

Ibuprophen                 PVOh                   800             8.1 x 10-4         54.63                    18

Fluoxetine                   PVOh                   20              1.2 x 10-5         54.37                    15

Tamoxifen                   PVOh                   20              1.1 x 10-5         47.61                      8

Ibuprophen                 PVOh                   300             3.0 x 10-4         47.43                    12

Aspirin                        PVOh              .0005M           5.0 x 10-4         45.57                    24

Fluoxetine                   PVOh              .0005M           5.0 x 10-4         44.92                    20

Vitamin C                    PVOh              .0005M           5.0 x 10-4         37.41                    14

Fluoxetine                   water                    60               3.6 x 10-5         37.31                      9

Aspirin                        PVOh                 325              3.8 x 10-4         36.02                    19

Acetomemophen         water                  1000             1.4 x 10-3         35.98                      4

Pure water                                                                        55               35.83                      3

Ibuprophen                 water                  800              8.1 x 10-4         33.66                      5

Fluoxetine                   water                    20              1.2 x 10-5         33.28                      2

Pure PVOh                                                                      48.9            32.05                      7

Aspirin                        PVOh                 2000             2.3 x 10-3         30.00                    25

Tamoxifen                   water                    20              1.1 x 10-5         29.05                      1

Aspirin                        water                    325             3.8 x 10-4         28.44                    6

Aspirin                        water                  2000             2.3 x 10-3         27.33                    13

Water index                                                                      55               26.84               Index

 

Background = 26.28

Clear Background = 66.06

 

Table 3: MRI Drug study – Scan set 3       

Scale: Structured =light = higher L value; Free = low L value

Compound                  Solvent            Dosage            Molarity         L Value            Position

Sucrose                        PVOh              .0005M           5.0 x 10-4         65.97                    16

Sucrose                        PVOh              .001M             1.0 x 10-3         65.82                    17

Celebrex                      PVOh              .001M             1.0 x 10-3         64.64                    15

Dextrose                      PVOh              .0005M           5.0 x 10-4         63.57                    18

Pure PVOh                  PVOh                                         48.9             58.75                    3

Caffeine                       PVOh              dose 200          2.2 x 10-4     57.00                    4

Pure Water                  water                                            55               56.80                    10

Phenobarbital  water   .001M                                    1.0 x 10-3         55.91                      9

Phenobarbital  PVOh  .001M             1.0 x 10-3                                 54.65                      2

Celebrex                      PVOh              .0005M           5.0 x 10-4         54.65                    14

Caffeine                       water               dose 200          2.2 x 10-4         54.44                    11

Caffeine                       PVOh              .0005M           5.0 x 10-4         52.40                      5

Dextrose                      PVOh              .001M             1.0 x 10-3         48.28                    19

Phenobarbital  water   .0005M                               5.0 x 10-4        48.08                      8

Celebrex                      water               .001M             1.0 x 10-3         45.07                    21

Glycerine                    PVOh              .0005M           5.0 x 10-4         44.54                      7

Sucrose                        water               .001M            1.0 x 10-3         44.02                    22

Caffeine                       water               .0005M           5.0 x 10-4         42.52                    12

Sucrose                        water               .0005M           5.0 x 10-4         42.35                    23

Phenobarbital  PVOh  .0005M                                  5.0 x 10-4        42.32                      1

Caffeine                       PVOh              .001M             1.0 x 10-3         39.39                    6

Celebrex                      water               .0005M           5.0 x 10-4         37.34                    20

Dextrose                      water               .0005M           5.0 x 10-4         35.00                    24

Caffeine                       water               .001M             1.0 x 10-3         32.04                    13

Water                          water                                            55               30.21               Index

Dextrose                      water               .001M             1.0 x 10-3         28.84                    25

 

Background = 26.01               Clear Background = 66.06

 

Table 4: MRI Drug study – Scan set 4       

Scale: Structured =light = higher L value; Free = low L value

Compound                  Solvent            Dosage            Molarity         L Value            Position

Pure PVOh                  PVOh                                           48.9           65.03                    3

Progesterone               PVOh              .0005M           5.0 x 10-4         64.52                    21

Cytoxan                      PVOh              dose 50            3.8 x 10-5         64.44                    11

Cytoxan                      PVOh              .0005M           5.0 x 10-4         63.26                    17

Pure PVOh-glass         PVOh                                           48.9            58.38                      2

Progesterone               PVOh              dose 10            5.8 x 10-6         57.15                    16

Imuran                         PVOh              .0005M           5.0 x 10-4         53.78                    18

Hydrocortisone           PVOh              dose 10            8.7 x 10-6         52.95                    15

Pure PVOh-glass         PVOh                                         48.9             51.89                    24

Imuran                         PVOh              dose 50            3.8 x 10-5         49.44                    12

Hydrocortisone           PVOh              .0005M           5.0 x 10-4         49.39                    20

Vioxx                           PVOh              dose 50            3.3 x 10-5         42.76                    8

Pure water-glass          water                                            55               38.53                    23

Pure water                   water                                            55               38.52                    22

Cytoxan                      water               dose 50            3.8 x 10-5         37.78                    4

Fructose                      PVOh              .0005M           5.0 x 10-4         37.62                    19

Pure PVOh-glass         PVOh                                         48.9             37.06                    6

Progesterone               water               dose 10            5.8 x 10-6         36.55                    10

Vioxx                           PVOh              .0005M           5.0 x 10-4         35.10                    14

Imuran                         water               dose 50            3.8 x 10-5         34.54                      5

Hydrocortisone           water               dose 15            8.7 x 10-6         34.75                    9

Fructose                      PVOh              .001M             1.0 x 10-3         33.08                    13

Pure PVOh-glass         PVOh                                           48.9            31.33                      7

Vioxx                          water               dose 50            3.3 x 10-5         28.43                      1

Fructose                      water               .001M             1.0 x 10-3              27.71                  25

Water                          water                                            55              26.75               Index

 

Background = 25.39                  Clear background = 66.06

 

Table 5: MRI Drug study – Combined Scan sets 1 – 4   

Scale: Structured =light = higher L value; Free = low L value

Compound                  Solvent            Dosage        Molarity         L Value   Position Scan

Tartaric acid                PVOh              1000                                       66.46               9         1

Caffeine                       PVOh              200                                        66.37               21        1

Vitamin C                    PVOh              250                                        66.22               11        1

Vitamin C                    PVOh                 60                                        66.11               4         1

Vitamin C                    PVOh              500                                        66.06               17        1

Sucrose                        PVOh              .0005M           5.0 x 10-4         65.97               16        3

Sucrose                        PVOh              .001M             1.0 x 10-3         65.82               17        3

Vitamin C                    PVOh              1000                                       65.04               23        1

Pure PVOh                  PVOh                                         48.9             65.03               3         4

Citric acid                    PVOh              .0005M           5.0 x 10-4         65.02               10        2

Tartaric acid                PVOh              500                                        64.97               2         1

Celebrex                      PVOh              .001M             1.0 x 10-3         64.64               15        3

Progesterone               PVOh              .0005M           5.0 x 10-4         64.52               21        4

Cytoxan                      PVOh              dose 50                                    64.44               11        4

Acetomenophen          PVOh                   650                                     64.43               11        2

Acetomenophen          PVOh                 1000                                     64.13               17        2

Caffeine                       PVOh                 50                                        64.01               15        1

Caffeine                       PVOh              .0005M           5.0 x 10-4         63.92               16        2

Dextrose                      PVOh              .0005M           5.0 x 10-4         63.57               18        3

Cytoxan                      PVOh              .0005M           5.0 x 10-4         63.26               17        4

Fluoxetine                   PVOh                   60                                      62.89               21        2

Acetomenophen          PVOh              .0005M           5.0 x 10-4         62.39               22        2

Ibuprophen                 PVOh              .0005M           5.0 x 10-4         62.21               23        2

Pure PVOh                  PVOh                                         48.9             58.75               3         3

Pure PVOh-glass         PVOh                                         48.9             58.38               2        4

Progesterone               PVOh              dose 10                                    57.15               16        4

Caffeine                       PVOh              dose 200                                  57.00               4         3

Pure Water                  water                                            55               56.80               10        3

Phenobarbital              water               .001M         1.0 x 10-3         55.91               9         3

Citric acid                    PVOh                 500                                       55.23               19        1

Phenobarbital              PVOh              .001M           1.0 x 10-3         54.65               2         3

Celebrex                      PVOh              .0005M          5.0 x 10-4         54.65               14        3

Ibuprophen                 PVOh                   800                                     54.63               18        2

Caffeine                       water               dose 200                                  54.44               11        3

Fluoxetine                   PVOh                   20                                      54.37               15        2

Imuran                         PVOh              .0005M      5.0 x 10-4         53.78               18        4

Citric acid                    PVOh                 60                                        53.39               6         1

Hydrocortisone           PVOh              dose 10                                    52.95               15        4

Caffeine                       PVOh              .0005M           5.0 x 10-4         52.40               5         3

Vitamin C                    water            1000                                        52.36               22        1

Pure PVOh-glass         PVOh                                         48.9             51.89               24        4

Citric acid                    PVOh              1000                                       49.86               25        1

Citric acid                    water               500                                        49.51               18        1

Imuran                         PVOh              dose 50                                    49.44               12        4

Hydrocortisone           PVOh              .0005M       5.0 x 10-4         49.39               20        4

Vitamin C                    water               500                                         49.08               16        1

Dextrose                      PVOh              .001M           1.0 x 10-3         48.28               19        3

Phenobarbital              water               .0005M       5.0 x 10-4         48.08               8         3

Vitamin C                    water                  60                                         48.03               3         1

Vitamin C                    water               250                                         47.83               10        1

Tamoxifen                   PVOh                   20                                      47.61               8         2

Ibuprophen                 PVOh                   300                                     47.43               12        2

Citric acid                    water               1000                                        46.63               24        1

Citric acid                    water               60                                          45.82               5         1

Aspirin                        PVOh              .0005M        5.0 x 10-4         45.57               24        2

Citric acid                    PVOh              250                                         45.51               13        1

Celebrex                      water               .001M         1.0 x 10-3         45.07               21        3

Fluoxetine                   PVOh              .0005M           5.0 x 10-4         44.92               20        2

Glycerine                    PVOh              .0005M           5.0 x 10-4         44.54               7         3

Sucrose                        water               .001M            1.0 x 10-3         44.02               22        3

Pure water                                                                       55               43.45               12        1

Vioxx                           PVOh              dose 50                                    42.76               8         4

Caffeine                       water               .0005M           5.0 x 10-4         42.52               12        3

Sucrose                        water               .0005M           5.0 x 10-4         42.35               23        3

Phenobarbital              PVOh              .0005M           5.0 x 10-4         42.32               1         3

Caffeine                       water               200                                         40.70               20        1

Caffeine                       PVOh              .001M             1.0 x 10-3         39.39               6         3

Pure water-glass          water                                            55               38.53               23        4

Pure water                   water                                          55               38.52               22        4

Pure PVOh                                                                     48.9             37.87               7         1

Cytoxan                      water               dose 50                                    37.78               4         4

Fructose                      PVOh              .0005M           5.0 x 10-4         37.62               19        4

Vitamin C                    PVOh              .0005M           5.0 x 10-4         37.41               14        2

Celebrex                      water               .0005M           5.0 x 10-4         37.34               20        3

Fluoxetine                   water                   60                                       37.31               9         2

Pure PVOh-glass         PVOh                                         48.9             37.06               6         4

Tartaric acid                water               1000                                        36.86               8         1

Progesterone               water               dose 10                                    36.55               10        4

Aspirin                        PVOh                 325                                      36.02               19        2

Acetomemophen         water                  1000                                     35.98               4         2

Pure water                                                                                         35.83               3         2

Vioxx                           PVOh              .0005M           5.0 x 10-4         35.10               14        4

Dextrose                      water               .0005M           5.0 x 10-4         35.00               24        3

Imuran                         water               dose 50                                    34.54               5        4

Hydrocortisone           water               dose 10                                    34.75               9         4

Ibuprophen                 water                  800                                      33.66               5         2

Caffeine                       water                    50                                      33.32               14        1

Fluoxetine                   water                    20                                      33.28               2         2

Tartaric acid                water                  500                                      33.13               1         1

Fructose                      PVOh              dose 50                                    33.08               13        4

Pure PVOh                                                                    48.9             32.05               7         2

Caffeine                       water               .001M        1.0 x 10-3         32.04               13        3

Pure PVOh-glass         PVOh                                         48.9             31.33               7        4

Water index                                                                      55               30.21               Index   3

Aspirin                        PVOh                 2000                                     30.00               25        2

Water index                                                                      55               29.49               Index   1

Tamoxifen                   water                    20                                      29.05               1         2

Dextrose                      water               .001M             1.0 x 10-3         28.84               25        3

Aspirin                        water                    325                                     28.44               6         2

Vioxx                          water               dose 50                                    28.43               1        4

Fructose                      water               dose 50                                    27.71               25        4

Aspirin                        water                  2000                                     27.33               13        2

Water index                 water                                            55               26.84               Index   2

Water  index                water                                            55               26.75               Index   4