R. SHEPARD, E. MCGOWAN, J. SCOTT, H, BAKER, D. BOWEN AND K. PRUITT 
585 
Table II. — Cerebrospinal Fluid Proteins of Calves Measured Before and After Total Cardiopulmonary Bypass 
Circulation of U Hours Duration with One-Third Hemodilution Prime 
After cardiopulmonary bypass 
Before 
Operation 2 hours 48 hours 3-11 weeks 
Total Protein (mg/100 ml) : 30 ± 11 64 ± 51 78 ± 22 98 ±65 
Albumin* 44.5 ± 8.4 52.0 ± 7.2 53.9 ± 4.7 39.4 ± 0.6 
Alphai-globulin 14.0 ± 5.1 8.7 ± 2.3 9.3 ± 3.0 12.4 ± 2.9 
Alphaa-globulin 11.3 ± 4.0 10.6 ± 2.4 9.9 ± 0.7 10.7 ± 1.8 
Beta-globulin 17.6 ± 4.7 15.4 ± 2.7 14.3 ± 2.7 14.0 ± 0.1 
Gamma-globulin 12.8 ± 5.1 13.3 ± 4.1 12.5 ± 1.6 23.4 ± 0.4 
Number of Animals _ 16 9 5 2 
♦Protein fractions determined by cellulose acetate electrophoresis and expressed as a percent of the total protein. Values expressed as 
mean ± standard deviation. 
Table III. — Serum Proteins Before and After Total 
Cardiopulmonary Bypass Circulation of ^ Hours 
Duration with One-Third Hemodilution Prime. Paired 
Data from 10 Calves 
Before 2 hours 
Serum Operation after bypass 
Total Protein (gm/100 ml) 6.6 ± 0.4 5.3 ± 0.8* 
Albumin, percent 51.3 ± 2.7 47.4 ± 3.3* 
Alpha-globulin 15.8 ± 1.9 13.7 ± 1.1* 
Beta-globulin 14.8 ± 1.9 15.5 ± 4.7 
Gamma-globulin 18.1 ± 3.0 23.4 ± 4.8* 
Protein fractions determined by cellulose acetate electrophoresis 
and expressed as a percent of the total protein. Values expressed 
as mean ± standard deviation. 
* p < 0.02. 
Table IV. — Calf Cerebrospinal Fluid Proteins Before 
and After Total Cardiopulmonary Bypass Circulation 
of i Hours Duration with One-Third Hemodilution 
Prime. Paired Data from 9 Calves. 
Cerebrospinal Before 2 hours 
Fluid Operation after bypass 
Total Protein (mg/100 ml) 35.1 ± 10.6 64.1 ± 53.7 
Albumin, percent 43.0 ± 10.8 52.0 ± 7.6* 
Alphai-globuIin 11.9 ± 2.9 8.7 ± 2.4* 
Alphas-globulin 13.0 ± 4.0 10.6 ± 1.5 
Beta-globulin 18.7 ± 5.3 15.4 ± 1.9 
Gamma-globulin 13.4 ± 5.1 13.3 ± 4.4 
Protein fractions determined by cellulose acetate electrophoresis 
and expressed as a percent of the total protein. Values expressed 
as mean ± standard deviation. 
* p < 0.05. 
Methods, Study 2 
Purified gamma globulin and albumin were 
used.* Solutions were prepared in buffer of 
pH 7.4, total phosphate concentration 0.004 M, 
and sodium chloride concentration 0.14 M. 
Gamma globulin solutions were centrifuged for 
twenty minutes at 27,000 g in order to remove 
insoluble material before being placed in the 
oxygenator. 
An otherwise standard disc oxygenator with 
* Obtained from the Pentex Corporation. 
its length shortened to 4 inches was used. The 
protein solutions were at room temperature, 
23-26°C. Room air was in the gas section of the 
oxygenator; it was not circulated. The number 
of revolutions of the discs was counted electro- 
mechanically. Total interfacial gas-liquid sur- 
face area produced during a given time was 
calculated using the area of the discs which 
became immersed in liquid during each revolu- 
tion, and the total number of revolutions during 
the time period. 
Relative turbidity was measured in a Brice- 
Phoenix light scattering photometer model 2000 
held at 25 °C. The light source was a mercury 
lamp fitted with a monochromatic filter with a 
436 m/x peak. Intensity of scattered light was 
measured at 90°. 
Fluorescence spectra were recorded with an 
Aminco-Bowman spectrophotofluorometer. The 
fluorescent probe l-anilinonapthalene-8-sulfon- 
ate (ANS) was purified by recrystallization 
and was used for the detection of albumin in 
the presence of gamma globulin.'^ Extrinsic 
fluorescence was measured by mixing an excess 
of the dye with protein solutions and measuring 
emission at 465 ± 10 m/ji (excitation at 375 ± 
10 mfji). 
Results, Study 2 
Gamma globulin solution turbidity develop- 
ment was nearly linearly related to the total 
interfacial area exposure, up to the point of 
precipitation of the denatured protein. The 
turbidity which developed was a direct function 
of the initial gamma globulin concentration. The 
maximum relative turbidity was 25 times higher 
than the initial value. When albumin, 1.6 mg/ml, 
was in solution vdth gamma globulin 0.4 mg/ml, 
