Michio Fujimaki

Preliminary evidence for inhibitory effect of glycyrrhizin on HIV replication in patients with AIDS.

Glycyrrhizin (GL) at a dose of 400-1600 mg/day (7.2-30.8 mg/kg/day) was administered intravenously for a period of more than a month, on 6 separate occasions, to 3 hemophiliacs with acquired immune deficiency syndrome (AIDS). Human immunodeficiency virus type 1 (HIV-1) p24 antigen was detected at the beginning of 5 of the 6 treatment courses. Viral antigen was not detected at the end of or during 3 of the 5 treatment courses and decreased to a low level following the 2 other courses. These findings suggest that GL might inhibit HIV-1 replication in vivo.

A new rapid and simple assay for factor XIII activity using dansylcadaverine incorporation and gel filtration

A new sensitive, reproducible and simple assay for factor XIII activity was developed, in which fluorescent dansylcadaverine (DC) is incorporated into casein and DC-casein is separated from free DC by gel filtration. A good correlation was found between our method and Lorands. The main advantages of our method are: (1) our assay technique is simple with good reproducibility and takes only a couple of hours; (2) the detectable limit of factor XIII activity is about 1% of normal pool plasma, but, if necessary, can be lowered by increasing the sample amount and lengthening the reaction time.

Mild hemophilia A resulting from Arg-to-Leu substitution in exon 26 of the factor VIII gene

SummaryPolymerase chain reaction amplification and nucleotide sequencing were used to identify the molecular defect in a Japanese patient with mild hemophilia A and an alteration of a TaqI site in exon 26 of the factor VIII gene. The mutation was a G-to-T transversion in codon 2326 of the factor VIII gene resulting in an Arg-to-Leu substitution at amino acid 2307 of the protein. The mutation, which is not of the common CG-to-TG type, is at the same codon in which both nonsense and a different missense (Arg to Gln) have previously been observed.

Possible cross-linking of factor VIII related antigen to fibrin by factor XIII in delayed coagulation process

The significance of factor VIII related antigen ( VIIIR:AG ) in laboratory medicine has been made clear, but its functions in the coagulation process are still unknown ( 1, 2 ). We previously observed that the concentration of VIIIR:AG in serum was lower than that in plasma of severe hemophilia A patients without inhibitor ( 3, 4 ). In this communication, we report the mechanism of the blood coagulation. consumption of VIIIR:XG in the state of prolonged

MspI polymorphic site in intron 22 of the factor VIII gene in the Japanese population

SummaryA novel MspI DNA polymorphic site has been found in intron 22 of the human factor VIII gene. This site is informative almost exclusively in the Japanese population (heterozygosity 0.45) and will be of considerable importance in carrier detection and prenatal diagnosis of hemophilia A in this population.

HIV markers and immune abnormalities in patients with hemophilia.

Etiological and immunological evaluations of 101 hemophiliacs treated with coagulation factor concentrates were performed. All samples were assayed for HIV antigen (Ag) with two different enzyme immunoassays (ABBOTT/VIRONOSTIKA), confirmed for antibody (Ab) to HIV with two different methods (ABBOTT ENVACOR/BIO-RAD westernblot). Results of serum HIV related markers assayed by these methods classify the patients into three groups: Of 101 patients, 37 in group A (Ab negative), 53 in group B (Ab positive but Ag negative), 11 in group C (Ab and Ag positive).OKT4 (helper/suppressor inducer) cells and OKT4/OKT8 (suppressor/cytotoxic) ratios in group C (323±164/μl and 0.48±0.23) were significantly lower than in group B (569±278/μl and 0.63±0.34) and A (737±367/4μl and 1.11±0.63).Among immunoglobulins quantified, serum levels of IgG and A were elevated in group C (2364±750mg/dl and 341±220mg/dl and 257±148mg/dl). Particularly, the inverse correlation between OKT4 cells and IgA levels was shown in HIV seropositive patients. The coefficient of correlation in group C (r=-0.341) was stronger than that in group B (r=-0.105). On the other hand, no relationship was found between IgM levels and seropositivity.Elevated serum β2-microglobulin levels were significantly correlated with reduced OKT4 cells in seropositive patients, especially in group C (r=-0.513).Our data suggest that assays of serum IgA, β2-microglobulin and HIV antigen are useful predictors for the condition and prognosis of HIV infection, furthermore the evaluation of therapy.

The replacement therapy with quantitative infusion of fVIII concentrate and its merit on plasma level and total dose.

On the basis of the in vitro stability of factor VIII concentrate at room temperature, continuous intravenous infusion of the remedey could be utilized as a hemostatic management for a major surgey in hemophilia A. One hour prior to the amputation of the left femule of the patient, 55u/kg of Hemofil H® as a bolus pretreatment and thereafter continuous infusion by quantitative pump with 2.7u/kg/hr of Hemofil H® for 3 days, 2.2u/kg/hr for next 5 days, 1.6u/kg/hr for the third period of 6 days, and 0.8u/kg/hr for last period of 4 days were followed. Consequently it was able to stabilize the hemostatic level of plasma VIII:C with the preparation, of which amount was less than that of previous cases with major surgical operations managed by bolus infusions of the similar preparation.Thus, this continuous infusion method applied on major surgery of hemophiliacs might be recommended as a thrapeutic way being safer and more economic than the conventional method, because no excess amount of factor VIII concentrate are infused.

Studies on α2-plasmin inhibitor and UK inhibitor in pregnancy and labor

To find out whether the depressed fibrinolytic activity in pregnancy can be ascribed to an increased content of inhibitors in the blood or to a release of inhibitors from the placenta, we studied the variation of the fibrinolytic inhibitors such as α1-AT, α2-MG, AT-III and C1-inactivator during pregnancy. However, we did not find any significant increase in inhibitors. In this paper, the levels of antiplasmin including α2-PI devised by Aoki and UK inhibitors extracted from the placenta by Kawano during pregnancy and delivery were examined.Methods:The material consisted of citrated plasma from pregnant women and extracts from the placenta. α2-PI was assessed with a single radial immunodiffusion method using antiserum made by Aoki. Antiplasmin activity was measured with chromogenic substrate S-2251 and then UK inhibitor was calculated from a standard curve prepared from placental UKI using the chromogenic substrate S-2444. The results were expressed relative to the content of normal standard as %, iu/ml and u/g.Results:1) The levels of α2-PI with a single radial immunodiffusion method correlated well to those of antiplasmin using the chromogenic substrate S-2251.2) Antiplasmin activity including α2-PI remained unchanged throughout pregnancy within 80-130%, and followed a similar patern after delivery. How-ever, it relatively decreased in retroplacental blood as well as uterine venous blood.3) In extracts of the placenta the concentration of antiplasmin was low.4) On the other hand, extract of the placenta contained a high concentration of UKI.5) The concentration of UKI was extremely low in each trimester, but the increased values were noted at term and in labor as well as in retroplacental blood, and gradually decreased after delivery.Discussion: Antiplasmin activity might directly or indirectly act upon the depression of the fibrinolytic activity in pregnancy, but UK inhibitor might be found intensively during placental separation.