The Mechanism of Hemolysis Caused by Uhrasonic Irradiation. I 

 W. RoMANOWSKi, A. Feltynowski, and J. Liiwin 



Human Physiology Depaitnient of the Academy of Medicine, Warsaw, 

 and State Institute of Hygiene, IVarsaw 



The authors investigated the mechanism of he- 

 molysis caused by the ultrasonic irradiation in several 

 experiments under special conditions, and they have 

 obtained some interesting results. 



The blood of the dog treated with hcparine and next 

 diluted three times by Tyrode's liquid was irradiated 

 for U min. from a generator of the power of 3 W/cm- 

 and at the frequency of 800 Kc. Then the specimen was 

 formed from the erythrocytes irradiated and from the 

 controls. The specimens were shadowcast with chromium 

 and were observed under the Siegbahn-Schoenander 

 electron miscroscope. 



All the blood cells from the irradiated specimen 

 were much smaller than the controls and attained ^ 

 of their previous size. The blood cells had no charac- 

 teristic "delta" form in the centre, and their shadows 

 were longer than those of the controls. This indicated 

 that the erythrocytes transformed into a spherical 

 form. 



We find that some results of Sibuya (2) and those 

 of Lindemann (1) though contrary to their inter- 

 pretation, give evidence that the spherical form is 



one of the forms in which the erythrocytes change 

 when subjected to ultrasonic treatment, and this 

 change may precede the hemolysis. It is now difficult 

 to decide whether it is caused by the denaturation 

 by the ultrasonic radiation of a characteristic protein 

 in the erythrocyte membrane which gives shape of a 

 "biscuit", or by the change in pH or temperature, 

 or by other factors. 



When irradiating with a smaller dose, changes 

 similar to those described by Lindemann were 

 observed, i.e. convex and concave deformations on 

 the surface of the blood cell. The appearance of these 

 forms can be explained by a non-uniform denatura- 

 tion of the protein over the whole surface. 



References 



1. Lindemann, B., Arcli. exptl. Put/iol. P/numakol. 209, 44 



(1950). 



2. Sibuya, after Bergmann, Der Ultraschall, p. 41. VDI- 



Verlag, Berlin, 1939. 



The Mechanism of Hemolysis Caused by Ultrasonic Irradiation. II 



W. RoMANOWSKi and A. Feltynowski 



Human Physiology Department of the Acadettiy of Medicine, Warsaw, 

 and State Institute of Hygiene, Warsaw 



By means of an ultrasonic 3 W/cm- generator at the 

 frequency of 800 Kc the authors irradiated mice in a 

 special vessel submerged into another one tilled with oil 

 (cooled with ice and water). The blood specimens were 

 taken from the tail and then investigated under the 

 electron microscope. The irradiation times were 9-1 3 min. 

 (mortal dosage) and 5 min. 



After irradiating with a mortal dosage nearly 

 all the erythrocytes were greatly changed. Their 

 sizes were much smaller than those of the control 

 ones, and their shapes were also changed. Some 



smaller or greater convex and concave deformations 

 were observed. After five minutes* irradiation such 

 changes have also been observed, but not on all 

 blood cells. 



These deformations are thought to be caused by 

 a non-uniform denaturation of the protein on the 

 surface, which gives a "biscuit" form to the erythro- 

 cytes. 



Similar changes in size and shape of the erythro- 

 cytes were obtained in /// vitro experiments (preceding 

 communication). 



1 1 — 568204 Electron Microscopy 



