Kaneo Yamada

Influence of granulocyte elastase-like proteinase (ELP) on platelet functions

The influence of granulocyte elastase-like proteinase (ELP) on platelet functions was investigated. ELP inhibited the platelet aggregations induced by a wide variety of agonists. The inhibition was marked in the case of receptor-mediated agonists such as thrombin, ristocetin, etc. It was moderate with the pervading agonist, arachidonic acid, and mild with the bypassing agonist, Ca2+ ionophore A23187. ELP inhibited the release of thromboxane A2 from platelets in the case of the platelet aggregation induced by thrombin. On the other hand, ELP did not inhibit the release of thromboxane A2 from platelets in the platelet aggregation induced by arachidonic acid or Ca2+ ionophore A23187. ELP suppressed the release of serotonin from platelets induced by thrombin, while it did not markedly suppress the release of serotonin induced by Ca2+ ionophore A23187. Treatment of platelets with ELP resulted in a slight increase of intraplatelet cAMP levels. These results suggest that ELP acts on receptors and inhibits platelet functions. As a results, ELP markedly inhibits the platelet functions such as aggregation or release of serotonin or thromboxane A2 stimulated by receptor-mediated agonists. ELP slightly elevates the cAMP level in the platelets, resulting in the mild inhibition of the platelet functions stimulated by the pervading agonist, arachidonic acid, or the bypassing agonist, Ca2+ ionophore A23187.

Three new electrophoretically normal glucose-6-phosphate dehydrogenase variants associated with congenital nonspherocytic hemolytic anemia found in Japan: G6PD Ogikubo, Yokohama, and Akita.

SummaryThree new glucose-6-phosphate dehydrogenase (G6PD) variants, which showed electrophoretically normal mobility and were associated with chronic nonspherocytic hemolytic anemia, were found in Japan. G6PD Ogikubo, found in a 17-year-old male whose red cells contained 3% of normal enzyme activity, had normal Km G6P, normal Km NADP, normal utilization of deamino-NADP, decreased heat stability, and a normal pH curve. G6PD Yokohama, characterized from a 15-year-old male, had 1.9% of normal enzyme activity, normal Km G6P, normal Km NADP, low Ki NADPH, normal utilizations of both 2-deoxy-G6P and deamino-NADP, decreased heat stability, and normal pH curve. G6PD Akita, characterized from a 56-year-old male, had an undetectably low activity when hemolysate was examined, normal Km G6P, normal Km NADP, normal Ki NADPH, normal utilizations of both 2-deoxy-G6P and deamino-NADP, decreased heat stability, and normal pH curve.The degree of hemolytic anemia was moderate to mild in all three patients.

A new APTT assay employing a chromogenic substrate and a centrifugal autoanalyzer

Abstract Using the synthetic chromogenic substrate for thrombin, S-2238, a new assay method based on the activated partial thromboplastin test (APTT) was devised with an autoanalyzer, Centrifichem system-400. Determinations at maximum optical absorbance of the reaction mixture and at half maximum, revealed good correlations between the values of the present APTT and those of the standard APTT: the correlation coefficients were 0.921 (p

A new assay method for factor XIII using a fluorescence polarization analyzer, based on change in the rotary brownian motion.

Abstract Employing a technique which determined the rotary Brownian motion by fluorescence polarization, an attempt was made to assay for factor XIII. The principle of the method was briefly as follows. Monodansyl cadaverine was converted to monodansyl casein by the action of factor XIII, so that the weight of the molecules with fluorescence was considerably altered; this then resulted in a lower rotary Brownian motion. If the alteration in level of rotary Brownian motion could be measured with a fluorescence polarization analyzer during the course of amine incorporation, the system should theoretically provide an assay procedure for factor XIII activity. By this method, a linear relationship between the alteration in fluorescence polarization and concentration of factor XIII or diluted normal plasma was obtained. A comparison was also made between the assay levels obtained by the present method and by the method by Lorand et al. The specificity of the present method for factor XIII was confirmed by inactivation of factor XIII with antihuman factor XIII A rabbit serum. For clinical purposes, the present assay method requires only a small amount of plasma (0.02μl) compared to the 1 μl needed in the previous fluorescence method. Also, it does not require different standard curves for different levels of factor XIII in the plasma, in contrast to the previous method. Another possible advantage is that the kinetics of the activity of factor XIII may be revealed with the assay procedure, although this point requires further assessment.

The effects of an alpha blocking agent on the respiratory distress syndrome with disseminated intravascular coagulation in the newborn infants and experimental animals

Abstract Firstly, the preventive effects of the alpha adrenergic receptor site blocking agent, phenoxybenzamine (POB) on rabbits which had been placed in an asphyxic condition, were observed. In the case of the asphyxiated rabbit which was treated with POB, shortening of recalcification time, and decrease in fibrinogen and platelets which were shown remarkably in the asphyxiated rabbit without POB, were clearly prevented. Secondly, the clinical trials of POB for the newborn infants suffering from RDS with DIC were carried out. One mg per kg of POB, was infused every 24 hours with alkali therapy and mechanical respiration. After the administration of POB, the clinical conditions improved as did the findings of coagulation studies. Little is known about the effect of alpha blocking agents on DIC in newborn infant with RDS.

Treatment of Childhood Acute Lymphoblastic Leukemia: the Results of the Tokyo Children’s Cancer Study Group L84-11 Treatment Protocol Study

The Tokyo Children’s Cancer Study Group protocol (TCCSG L84-11), a randomized trial, was designed to evaluate moderate dose methotrexate (MD-MTX) therapy plus late skull irradiation and intensified central nervous system (CNS) prophylaxis by triple intrathecal (IT) therapy. From 1984 to 1988, children with newly diagnosed acute lymphoblastic leukemia (ALL), except for B-ALL, were enrolled in this protocol. Patients at high risk had one or more of the following risk factors: age below 2 or above 7 years, white cell count of 20,000/μ1 or more. Patients at extremely high risk had the following risk factors: T-cell phenotype, white-cell count of 100,000/μ1 or more, or mediastinal mass. Other patients were in the standard risk group. The standard risk (S) regimen consisted of two arms (S-1, S-2). For the remission induction, conventional VPL therapy was carried out. After the remission, triple IT therapy and 18 Gy radiation were given to S-l, and MD-MTX, late radiation (24 weeks later) and maintenance therapy was the standard form. The therapy was terminated after 3.5 years. The high-risk (H) regimen consisted of two arms (H-l, H-2). The outline of this regimen was similar to that of the S regimen. In the H regimen, cyclophosphamide, anthracyclines, BH-AC, cytosine arabinoside, and 1-asparaginase were administered for cyclic intensification therapy. The extremely high risk regimen (Hex) was a modified protocol of BFM-ALL83. There were 471 eligible patients. The remission rates were 97–100%. The patients with relapsed disease were 9/93 (S-1), 8/92 (S-2), 21/127 (H-1), 20/113 (H-2), and 8/46 (Hex). Event = free survival rates (EFS) were 80%, 66-74% and 69% in the S-, H-, and Hex regimens, respectively. In each group, no significant difference in EFS was demonstrated between the two arms (S-l vs. S-2, and H-l vs. H-2). The most common site of relapse was the bone marrow (49/66, 74%). Patients with CNS relapse were 5/93 (S-1), 2/92 (S-2), 1/127 (H-1), 0/113 (H-2), and 2/46 (Hex). The unexpected sequela of this protocol was the severe neurotoxicity that accumulated in the H-2 subgroup. Although the follow-up period is short, these preliminary results indicate that this protocol apparently reduces the CNS relapse rate, especially in the high-risk group. However, further modification of CNS prophylaxis is necessary to avoid neurological sequela.