AD-Ace5 223 TEXAS INST FOR REHABILITATION AND RESEARCH HOUSTON F/6 14/2NAVY ENVIRONMENTI DIELECTRIC STUDIES OF BIOLOGICAL NATERIALS. (U)MAY 80 0 S KIRTPATRICK N00014 79-C-0076

UNCLASSIFIED TRI1 NL

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MICROCOPY RESOLUTION TEST CH-RTNATIONAL BUREAU OF STANDARD6-1963-l

OFFICE OF NAVAL RESEARCH

Contract N00014-79-C-0076' ".

Task No. NR 207-176

i-

Technical Report No. I

Navy Environment: Dielectric Studies

00 of Biological Materials

0 byDonald S. Kirkpatrick 10

Instw e-for Rehabilitation and Research/1333 Moursund Avenue

Houston, Texas 77030

30 May 1980

Reproduction in whole or in part is permitted forany purpose of the United States Government.

Distribution of this report is unlimited.

80 6 5 030I ____ _ - -

I

ONR. I FINAL REPORT

Technical Report No.1

Navy Environment: Dielectric Studies of Biological Materials

Our accomplishments under this contract include the following topics:

1. Data acquisition from time-domain reflectometer

2. Preparation of. manuscript describing our preliminary dielectricspectroscopy of membrane suspensions

3. Preparation of manuscript for research proposal on dielectricspectroscopy of water-macromolecule interactions

4. Rheological investigation of cystic fibrosis mucus and preparation ofmanuscript

Each of these topics is discussed below.

1. Data Acquisition from Time-Domain Reflectometer

Data acquisition from the time-domain reflectometer by microcomputer becamefirst priority in this project due to prolonged failure of our original dataacquisition computer.

a. Selection of appropriate microcomputer

We selected a data acquisition computer to satisfy several criteria. Werequired more than 16K bytes of internal memory in order to accomodatesimultaneously 2000 to 4000 data points (two bytes each) from each of two scans,viz., sample and reference scans. Sufficient external memory was needed tostore scan pairs from multiple experiments. For this task we selected aCromemco System II microcomputer with 32K bytes of random access memory and two5 inch minifloppy disc drives. This instrument also provided both BASIC andFORTRAN IV for ease in computations as well as an assembler, for dataacquisition.

b. Time-domain reflectometer-computer interface

The data from the sampling oscilloscope consist of analog signals from itshorizontal and vertical output terminals. These outputs represent the X-Ycoordinates of the trace displayed on its screen. The sampling oscilloscopealso generates from its pre-trigger output 3000 regularly spaced pulses pertrace.

Our task was to provide the logic interface between the horizontal andpre-trigger outputs of the oscilloscope and the digital interface of thecomputer. This interface uses the pre-trigger output pulses from theoscilloscope to command acquisitition of each data point in the trace, and the

discontinuity of the horizontal ramp signal to initiate and terminate each

trace. The data are acquired from the vertical output of the oscilloscope using

a sample/hold amplifier to sample the analog signal followed by ananalog/digital converter to digitize the sampled value. The computer then

stores the value in memory. The logic interface allows program control of the

data acquisition. There was no attempt to control the output of the oscilloscope

by the computer.

The schematic diagram of the logic interface board is shown in following

diagram along with the the timing diagram and flow' chart for. the data

acquisition program.

2. Preparation of Manuscript: HYDRATION DEPENDENT, COLLECTIVE DIELECTRICBEHAVIOR OF ROD OUTER SEGMENTS

ONR.2We prepared a manuscript which reports our preliminary data on the

dielectric behavior of membrane systems. This manuscript is summarized -asfollows.

SUMMARY

We observed in concentrated aqueous suspensions of rod outer seiments a"slow" dielectric process with a relaxation time of about 10W . The

"slow" process was about 103 slower than the dielectric relaxation time ofsolvent water. The dielectric constant of the "slow" process 1.) variedwith the hydration level of the suspension, reaching a maximum greater than1000 in the hydration range which equaled the water content reported forrod outer segments in vivo, 2. ) correlated with conductivity and ionicstrength of the suspending solution, and 3.) was too large to be explainedby linear combinations of water and membrane, which display dielectricconstants of 78 and 2, respectively.

The following causes for the "slow" dielectric process were inconsistentwith our observations: 1 .) dipolar rotations of proteins in the plane ofthe membrane, 2.) membrane gating currents, and 3.) interfacialpolarization, viz., Maxwell-Wagner and frequency dependent, transverseconductance and capacitance. We suggest a collective or many-bodyinteraction between water, ions, and membrane macromolecules to explain themagnitude of the dielectric constant, its dependence upon hydration, andits correlation with conductivity.

Copies of this manuscript have been submitted to the Program Director,..Biophysics.

3. Preparation of Manuscript for Research Proposal: INVESTIGATION OFWATER-MACROMOLECULE INTERACTIONS BY DIELECTRIC SPECTROSCOPY ONR.4

This manuscript 1.) summarized our dielectric experiments, 2.) presentedthe experimental details of the time-domain (Fourier transform) dielectricspectroscopy technique, 3. ) reviewed water-protein interactions, and 4.)presented a rationale for investigating water-protein interactions by dielectricspectroscopy. This manuscript has been submitted to the Program Director,Biophysics. It will be submitted to various foundations and institutes* as agrant proposal in the future.

4. Rheological Investigation of Mucus OmR.3

We correlated physical and chemical properties of mucus in order to

determine the chemical basis for mucus becoming obstructive in obstructive mucusdiseases such as cystic fibrosis. A manuscript, SALIVARY MUCUS: CORRELATIONBETWEEN FLOW PROPERTIES AND CHEMICAL PROPERTIES IN CYSTIC FIBROSIS SALIVA

reported this work, and has been submitted to the Program Director, Biophysics.It is summarized below.

SUMMARY

In order to determine the chemical basis for mucus becoming obstructive, wesimultaneously investigated both the rheological properties of mucus and itschemical composition. We estimated the value of Tow (tackiness or Spinnbarkeit)and quantitatively measured six chemical parameters of individual specimens ofsalivary mucus from cystic fibrosis patients. We found that we could describethe relationship between Tow and the chemical parameters by the multivariatoequation:

Tow = 2.75(Alb.) + 0.456(T.Pro.) + 0.024(Cond.) - 1.241(T.Carb.)+ 0.001(Amy.) + 0.355. (Equation 1)

Equation 1 predicts the value of Tow from the chemical constituents of themucus, viz., endogenous concentrations of albumin, total protein, inorganic ions(as measured by conductivity), total carbohydrate, and alpha-amylase. It issignificant at the p = 0.1 level, and accounts for 25% of the variabilityobserved in Tow (multiple R squared = 0.25). With more data points thisrelationship can become significant at the p = 0.05 level. Our data establishedfor the first time that rheological properties of CF salivary mucus correlatewith concentrations of its multiple chemical components.

These correlations suggest roles for albumin, mucin glycoproteins, andinorganic ions in enhancing, and for unidentified carbohydrate containingmaterial in diminishing, visco-elasticity of CF mucus. Equation I wasindependent of wide, week to week fluctuations, which we observed in both Towand chemical parameters of individuals. Such fluctuations precluded correlationof Tow with the presence or severity of CF disease. Tow fluctuations probablyresulted from the fluctuations which we observed in the concentrations ofchemical constituents, rather than from possible fluctuations in mucinglycoprotein structure or function, as previously suggested.

Our preliminary results confirm the effectiveness of our strategy forinvestigating obstructive mucus. This strategy consists of simultaneouslymeasuring both the rheological properties and the concentrations of multiplechemical constituents of individual specimens of CF salivary mucus. Theirrelationships are examined by multivariate analysis. On this basis we proposethree immediate goals:

1.) Verify our preliminary results using good statistical design.2.) Refine the original physical and chemical parameters of saliva to more

specific and well defined parameters.3.) Investigate the cause and effect relationships between rheological

properties and chemical constituents, which the correlations suggest

Following this strategy, we expect to determine how the theological

properties of mucus are physiologically regulated as well as how obstructivemucus can be managed clinically. We hope that this knowledge will enable us toprevent the fatal lung damage which CF inflicts.

7 DATA ACQUiIiT LON

-Logic Interface between Time-Domain Ref lectometer and

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ii!.DATA ACQUIS9ITIrONTiming Diagram between Time-Domain Ref lectometer and

TU-ART Digital Interface (Cromemco)

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Navy Environment: Dielectric Studies of Biologic Final Repert.Maeials,& Nov 10VW - Feb-9-, 198C

S. PERFORMING ORG. REPORT NUMBER

7. ATHOR~a .1. CONTRACT Oft GRANT NUMBER(&)

(6 Donald S.; Kirkpatrick 4,.E~W 9 * C

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The Institute for Rehabilitation and Research AE OKUI UUR

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DIELECSTRC (Coniu ntermsCos,ai reach proposal, 3.) SWeeAk IUCU:*O LA

reportsigerrhelogicl/ohewic indestato ofquisstic fibgrosiso acus.

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BIOLOGICAL SCIENCES DIVISIONBIOPHYSICS PROGRAM, Code 444

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