Richard F. Riley

Isolation of C-reactive proteins of man, monkey, rabbit and dog by affinity chromatography on phosphorylated cellulose.

Abstract The ability of C-reactive protein (CRP) to complex with organic monophosphates in the presence of calcium was exploited in devising a method for isolating this protein in a simple, highly selective way employing phosphorylated cellulose. The same procedure was found to provide CRPs from monkey, rabbit and dog as well and may prove to be a general procedure for the isolation of CRP of other species.

Heme synthesis and erythrocyte life span in the cat.

Summary The mean life span of the erythrocyte in the cat as determined by labeling heme with N15 is 77 days.

Effect of whole-body and partial-body x-irradiation on the extractable cellular components of the lung with special consideration to the alveolar macrophage.

Female Sprague-Dawley rats were given various whole-body and partial-body x-ray exposures to determine the radioresponsiveness of extractable cellular components of the lung. Cellular suspensions were obtained from the lung by intratracheal lavage with a balanced salt solution. Exposure of the thorax to 1000 rad and whole-body irradiation at three dose levels (500, 750, and 1000 rad) all reduced extractable alveolar macrophage numbers to approximately 50% of control values during the second week after irradiation. Recovery was indicated by the fourth week. The number of polymorphonuclear leucocytes extracted from the lung increased to approximately ten times control levels during the third post-irradiation week and subsequently decreased in an exponential manner. Small aphagocytic mononuclear cells were recovered from lungs of all control rats. Their numbers were reduced prior to the third week after irradiation. Mean macrophage diameters and the frequency of binuclear macrophages were increased following...

CX-REACTIVE PROTEIN RESPONSES IN THE RABBIT AFTER WHOLE-BODY IRRADIATION

Cx-protein was present in the serum of all rabbits exposed to whole-body doses of 250 to 750 r of x rays 24 hours after exposure, and the maximum Cx- protein titer was usually observed at this time. Not all animals gave a Cx- protein titer 24 hours after 100 r or less. The mean 24-hour titer was about the same after doses of 100 to 750 r. The response was commonly biphasic in fed rabbits and monophasic in rabbits denied food during the postirradiation period. Treatment with compound 48/80 produced a Cx-protein response and animals so treated behaved like controls when subsequently given 250 r. Pyrilamine did not modify the Cx-protein response to 250 r of x rays. (auth)

Behavior of the c-reactive protein during purification and electrophoretic characterization☆

Abstract C-reactive protein prepared chromatographically on DEAE cellulose is obtained in part as an aggregated complex with the mobility in 7 1 2 % polyacrylamide typical of 7 S γ-globulin. The aggregated protein slowly dissociates on chromatography and on electrophoresis in polyacrylamide gel to give components antigenically related to C-reactive protein with electrophoretic mobilities in polyacrylamide similar to those of pre and post albumins. This is also true for C-reactive protein prepared by the conventional method. The electrophoretic mobility of C-reactive protein in polyacrylamide depends on the preparative history and age of the sample. The relative mobilities of its subunits were independent of method of isolation, but differed in their mobility from one individual to another, suggesting the existence of multiple molecular forms of this protein. The polyacrylamide matrix in which proteins have been resolved electrophoretically can be conveniently adapted to provide immunoelectrophoretic data.

Cobalt Color Reaction of Barbiturates.

Of the numerous analytical methods employed for the determination of barbiturates, those making use of specific color reactions have been most extensively studied. Handorf 1 employed the mur-exide reaction for the detection of veronal, and Zwikker, 2 Bodendorf, 3 Herwick 4 and Koppanyi, et al., 5 , 6 have developed modifications of a color reaction with cobalt salts in conjunction with various bases. Kozelka and Tatum 7 attributed this color formation to a specific reaction with substances containing one or 2 imide groups. In an investigation of amytal (isoamylethyl barbituric acid) excretion by dogs, using a modification 9 of the reaction designed by Koppanyi et al., 6 employing cobalt acetate and isopropylamine made up in absolute methanol solutions, it was observed in this laboratory, that of the more probable breakdown products of amytal 8 only isoamyl-ethyl acetic acid gave the characteristic color reaction. Since this compound contains no imide grouping, it was apparent in this case that color formation was not due to such a group. This finding led to an examination of various classes of compounds a number of which have not been reported upon by other investigators. A partial listing is given in Table 1. Since the acids tested gave a color apparently identical with that obtained with amytal, a number of these test solutions were submitted to spectrophotometric analysis. The spectrophotometric curves given herewith were made with a Bausch and Lomb medium quartz spectrograph and sector photometer. The length of the absorption cells was 5 cm. To cobalt acetate and isopropylamine mixtures were added respectively, solutions of amytal, isoamyl ethyl acetic acid (RRC2H2O2) and stearic acid in amounts sufficient to give an intensity of color suitable for measurement.

Identification of Ferritin in Blood of Dogs Subjected to Radiation from an Atomic Detonation

Summary Studies on blood plasma samples obtained from eight dogs subjected to a lethal dose of radiation from an atomic detonation demonstrated that a material was present which, when administered intravenously, decreased the rate of vasomotion and decreased the epinephrine sensitivity of the mesoappendix capillary bed of the normal rat. After its appearance in the blood, the concentration of this vasodepressor material decreased as a function of time. This VDM has been shown to fit the criteria established for ferritin.

Effect of drugs on Cx-protein responses in the rabbit.

Summary Methyl prednisolone and fluorometholone prevented the appearance of CxRP in rabbit serum following subcutaneous injection of mineral oil-Aquaphor emulsion. Fluorometholone was unable to block CxRP responses to 250 r whole body irradiation or to intravenous injection of Thorotrast or Varidase. It did block CxRP responses to intravenous injection of post irradiation seromucoid or TMV and the intravascular formation of antigen-antibody complexes. Under appropriate time dose conditions fluorometholone inhibited both antibody production and CxRP appearance while under other time dose conditions CxRP appearance was prevented without alteration of antibody production.