A given quantity of cysteine injected intravenously in the normal rabbits produced two peaks in leucocyte count, but the second peak disappeared when the cervical cord was severed. In the rabbits with the liver injured by CCl4, cysteine showed same tendency as in the normal rabbits. There appeared, however, a marked first peak and a diminished low second peak. These facts show that the leucocyte increase occurs in two ways such as the peripheral first and the central second peak. Now with the purpose of clarifying these relations, investigations have been made as to which leucocyte-increasing peak Neutrophilin (Neutropoetin-Komiya) is proved in the serum-Neutrophilin which is inevitably produced in the serum at the time of the central leucocyte increase. The results are as follows: 1. The serum three hours after the administration of cysteine did not contain Neutrophilin. The first increasing peak (appeared three to four hours later), therefore, was caused peripheral-produced by the direct stimulation of the hematopoietic organs and it was not the “central” leucocyte increase. 2. The existence of Neutrophilin was proved in the serum 7 hours after the administration of cysteine. The second peak (occured 8 hours later), therefore, was the central leucocyte increase.
What is the relation between the leucocytosis in the peripheral blood caused by the intravenous injection of cysteine and the bone marrow cell? In order to make researches upon this problem, 6mg of cysteine was given intravenously to the normal rabbits and the bone marrow pictures were studied 4 hours and 8 hours after the injection-when the increase was observed in the leucocyte count in the peripheral blood-and also after 24 hours-when the count dropped back. The results are as follows: After 4 hours and 8 hours the nucleated cell count decreased markedly with the variations of the bone morrow picture primarily due to pseudoeosinophils, and especially mature cells diminished. while, the immature pseudoeosinophils were observed to have a tendency to increase to some extent. Considering the above results, the leucocytosis with characteristics two-peaks-mainly due to pseudoeosinophil increase by cysteine administration-is not resulted from the release of the reserved leucocytes from the blood cell reservoirs, but obviously from a fact that the leucocyts is mobilized out of the bone marrow. Further, the previous reports and literature lead us to infer that the first peak of leucocytosis, appearing 4 hours after the injection of cysteine, is formed by the direct action of cysteine itself to the bone marrow, and that the second peak occurring 8 hours after the injection is produced by secondarily produced Neutrophilin (Neutropoetin) acting to the bone marrow-both cysteine and neutrophilin (Neutropoetin) promotes hematopoiesis of pseudoeosinophils while inducing the mature pseudoeosinophils to be mobilized and to move out.
In my previous report, the transfer rate of albumin was studied by using albumin I131. In the present study, the transfer rates of body fluid in the various diseases were investigated by using radio phosphate (P32) and sodium thiocyanate (NaSCN), following the intravenous and intraperitoneal injection. The results were as follows; (1) The disappearance rate of P32 from the plasma in patients with ascites was greater than those in patients without ascites. (2) The transfer rate of P32 and NaSCN into ascites following the intravenous injection in carcinomatous peritonitis and tuberculous peritonitis were greater than those in cirrhosis of the liver and Banti's syndrome. (3) The transfer rate of P32 into plasm from ascites following intraperitoneally injected P32 was increased in cirrhosis of the liver and decreased in carcinomatous peritonitis. (4) The transfer rate of of P32 and NaSCN into the ascites was parallel to the clinical course of cirrhosis of the liver, Bantis syndrome and tuberculous peritonitis. It's rate was decreased according to the clinical remission. (5) The transfer rate of NaSCN was decreased with the decreasing ascites in the cases which had showed the significant effect by the use of mercurial diuretic and hydrocortisone. (6) The transfer rate of P32 and NaSCN was decreased after abdominocentesis following the intravenous injection. It's rate was increased after abdominocentesis following the intraperitoneal injection.
The author demonstrated in the previous report that the sera of patients with rheumatoid arthritis are able to agglutinate in high titers the human O erythrocytes sensitized by a non-agglutinating amount of rabbit anti-human O cell serum. This modified Waaler-Rose test was used in the present study and following results were obtained. 1. The sensitized human O cell agglutination reaction was significantly potentiated when patients sera were diluted in 5% sheep or guinea pig serum rather than in salein solution. When 5% horse, dog and human sera were used as diluents, the sensitized O cell agglutination titers were either somewhat increased or unchanged, whereas the agglutination was significantly inhibited when patients sera were diluted in rabbit serum. 2. Increase in titer more than fourfold was observed in 71% of 28 sera of patients with rheumatoid arthritis when diluted in sheep serum and in 75% of the same cases when diluted in guinea pig serum. On the other hand, the same increase in titer was observed in 12% of 42 sera of patients other than rheumatoid arthritis when diluted in sheep serum and in 14% when diluted in guinea pig serum. 3. The solutions of Cohn Fraction II+III (globulin) of sheep and guinea pig serum potentiated the sensitized human O cell agglutination reaction when sera of patients with rheumatoid arthritis were diluted in these solutions. 4. When solutions of various concentrations of human serum Cohn F.V, F.II, F.III and F.II+III were used as diluents, the sensitized human O cell agglutination titer increased in F.V solutions (especially in relatively low concentrations), and decreased in F.II, F.III and F.II+III solutions.
Presented in this paper are the results of some fundamental investigations on the technical aspects of the thromboplastin generation test, firstly described by Biggs and Douglas in 1953, and on the nature of the coagulation factors involved in the formation of an active prothrombin converting principle or blood-thromboplastin by employing thisva luable technique. For the present study, some modifications in the thromboplastin generation test as originally described are used: the use of BaSO4 instead of Al (OH)3-gel for the absorption of prothrombin, numerical standardization of the platelet suspension. 1) Five factors, platelets, anti-hemophilic globulin, plasma thromboplastin component, and factor V and VII in the presence of CaCl2, seemed to be necessary for blood-thromboplastin formation. 2) Experiments with brain extract have suggested that brain extract is not itself thromboplastic but can be converted to thromboplastin after interaction with factor V and serum fraction. 3) The amount of blood-thromboplastin generated is quantitatively related to the number of platelet, concentration of anti-hemophilic globulin. The greatest effect on the speed of blood-thromboplastin formation, on the other hand, is related to the concentration of anti-hemophilic globulin. 4) Platelet suspension may be kept for about a week without any significant loss of activity, when stored in refrigerator at-1°C. This makes it possible to run the thromboplastin generation test as a routine screening test. 5) The adsorbed plasma and the serum should be diluted at least half an hour before the test are performed, though the explanation of this phenomenon has not yet been made. 6) Relative lack of factor V in the reacting mixture has little influence on the thromboplastin generation test in the presence of fresh platelets. This may be due to platelet factor 1. 7) The activity of blood-thromboplastin formed in reacting mixture disappears rapidly. If a buffer is introduced in the reactives, a much more stable activity can be obtained. Anti-hemophilic globulin and factor V are consumed in the course of blood-thromboplastin formation. 8) Concentration of thrombin formed in the reacting mixture of the thromboplastin generation test is very low and is too small in amount to clot substrate plasma. Small amount of thrombin accelerates the speed of blood-thromboplastin formation in the original test, but no such effect has been observed in the presence of brain extract. It may be inferred, therefore, that the accelerating action of thrombin takes place at an initial stage of blood-thromboplastin formation.