Chapter 9, Note 2. Lipids And Rouleaux And Sludge Formation

Human blood obtained through venous puncture was mixed in the syringe with 1/10 of its volume of 1.5% sodium citrate, passed into several centrifuge tubes and separated into plasma and cells. The plasma from the various centrifuge tubes was placed in separate test tubes. One of these samples of plasma was treated with a mixture of conjugated fatty acids of cod liver oil, another with a lipoacid preparation of human placenta, a third with a preparation of the insaponifiable fraction of human placenta. As control, the plasma was treated with liquid paraffin. The mixtures, frequently shaken, were kept in a warm bath at 37°C for one hour, after which they were centrifuged and the oily material separated. The plasma treated with the lipoacids was then added in proportion of 10% to untreated plasmas which then were reunited with their red cells. The plasma treated with the insaponifiable fraction was mixed directly with its respective red cells. Plasma and red cells were shaken for five minutes and left at room temperature for another ten minutes. One small drop of this blood obtained with a platinum loop or a capillary pipette was mixed with two drops of saline on a slide covered by glass and examined under the microscope. While the controls showed few short rouleaux, the blood treated with insaponifiable fraction showed isolated cells. In the blood treated with placenta lipoacids, almost all of the red cells formed rouleaux; in blood treated with conjugated fatty acid, almost all cells formed sludges.

Chapter 9, Note 3. Dark Color Of The Blood In Shock

In order to determine why dark colored blood is seen in shock and the role of fatty acids, the following experiments were performed.

Venous blood of patients in severe state of shock was drawn and mixed with 1/10 of its volume of a citrate solution. The hematocrit value was determined and saline added in order to bring it to the value of the normal blood. Through these blood samples, oxygen was passed for five minutes at the rate of 50 cc. per minute. At the same time, blood from a normal person was obtained and similarly treated. The changes in color of the two samples after cessation of oxygenation were compared. While the normal blood needed almost ten minutes to return to the previous color, the blood from the patient in shock was back to the deep dark color in less than three minutes.

Blood samples from subjects in shock were treated in vitro with un saponifiable fractions and subsequently with oxygen for five minutes at the rate of 50 cc. per minute. While the color became red immediately, the time required for it to return to the previous dark color was entirely different from that of controls. As against a few minutes for controls, more than twelve minutes were required for the blood treated with insaponifiable fraction. From these experiments it appears that the dark color of blood in shock results from changes in the red cells and not because of impaired circulation, and that these changes can be related to the intervention of fatty acids. This was confirmed by the fact that the dark blood of shock patients, if treated in vitro with insaponifiable fractions of placenta, for instance, loses its characteristic color.

Chapter 9, Note 4. Induction Of Acute Shock

Acute shock can be induced by intraperitoneal injections of mixtures of conjugated fatty acids. The preparation largely used was a 10% solution in oil of fatty acid of cod liver oil conjugated through treatment with KOH in ethylene glycol or in ethyl alcohol. For a rat of 200 grams, 8 cc. of this preparation injected at once was able to induce an acute shock.

Chapter 9, Note 5. Induction Of State Of Shock

State of shock was induced by the repeated administration of a mixture of conjugated fatty acids obtained from cod liver oil. To insure a progressive systemic absorption, the preparation was injected subcutaneously. The injection in rats of 1 cc. per 100 gram of body weight of the 10% solution of these fatty acids in oil, repeated every hour was seen to induce after 3-5 injections a state of shock. The addition of 4% of sodium thiosulfate in a dose of 5 cc. per 100 gram of body weight in rats, was seen to favorize the appearance of this state of shock.

Chapter 9, Note 6. Influence Of Fatty Acids Upon Traumatic Shock

Rats of 250 grams were introduced in the Collip Noble Drum with their forepaws taped with adhesive and submitted to 500 falls at a rate of 40 per minute. 50% died of acute shock in less than two hours. If 2 cc. of a preparation of 10% cod liver oil fatty acids in oil per 100 grams of body weight was injected intraperitoneally or even subcutaneously 1/2hour before the animals were placed in the drum, more than 50% of the animals died during the trauma itself and the fatality rate in some experiments approached 100%. Bleeding from the nose and mouth was dark in color and smaller in quantity than in untreated animals. If the same amount of the lipoacid preparation was injected immediately after the animal came out of the drum, it also increased mortality within the first two hours. For some animals, death occurred in a few minutes after the injection.

Chapter 9, Note 7. Influence Of Unsaponifiable Fractions Upon Traumatic Shock

The influence exerted by the unsaponifiable fractions upon traumatic shock appeared evident in rats submitted to 5-700 falls in the Collip Noble drum. 10% solutions in sesame oil of the unsaponifiable fraction of human or cow placenta, of eggs or of butter, were used in these experiments. 1 to 5 cc. of these solutions were injected intraperitoneally at different intervals before or after trauma. The injection of 2 cc, 1/2 hour before trauma, was seen to entirely prevent lethal shock (0/20) in a group of experiments where the mortality of controls was 18/20. The same results were obtained with 2-3 cc. of the preparations injected immediately after the animals were taken out of the drum. Doses as high as 5 cc. injected one hour after the animal was removed from the drum, protected only a few animals (11 /20) and generally only those without symptoms of shock. Once the symptoms of shock were present, the effect of the unsaponifiable fractions was greatly reduced. (From 2/20 to 5/20 in different experiments.)