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Background One of the proposed aetiological mechanisms for idiopathic sudden deafness is vascular disease. However, it is not known whether traditional cardiovascular risk factors, such as particular dietary factors, are associated with this condition. Methods A case-control study using pooled controls was conducted in Japan to investigate the relationship between idiopathic sudden deafness and diet. An m:n matchedpairs method was used to obtain age-, gender- and residential district-matched controls from a nationwide database of pooled controls. Food intake was assessed from a self-administered usual food frequency questionnaire that asked about intake of 35 foods (including four drinks). Participants were classified according to the frequency of intake of Western foods and the frequency of intake of traditional Japanese foods. Subgroup analyses were performed using audiometric subtypes of idiopathic sudden deafness. Results Data were obtained for 164 cases and 20 313 controls. An increased risk of sudden deafness was observed among participants who frequently consumed Western foods (OR = 1.82, 95% CI : 1.14‐2.89), and a decreased risk of this condition was observed among participants who frequently consumed Japanese foods (OR = 0.52, 95% CI : 0.33‐0.82). A direct association of sudden deafness with Western food intake was evident for flat-type hearing loss. Conclusions This study suggests that a largely Western diet might be a risk factor for idiopathic sudden deafness, a traditional Japanese diet might be a preventive factor for this condition, or both. These findings are consistent with the hypothesis that vascular factors are an important cause of idiopathic sudden deafness, although the possibility of residual confounding by unmeasured confounders such as socioeconomic status cannot be ruled out.

Sudden deafness sometimes has an identifiable cause, but in most cases the cause is unknown (idiopathic sudden deafness). Vascular impairment has been proposed as an aetiological mechanism for this condition, but it is unclear whether traditional cardiovascular risk factors, such as smoking or alcohol intake, are associated with this condition. We accordingly investigated associations of idiopathic sudden deafness with smoking, alcohol intake and sleep duration in a case-control study. Cases were consecutive patients diagnosed with idiopathic sudden deafness between October 1996 and August 1998 at collaborating hospitals in Japan. Controls were obtained from a nationwide database of pooled controls, with matching for age, gender and residential district. Exposure variables were assessed from a self-administered questionnaire. Subgroup analyses were performed using audiometric subtypes of sudden deafness. Data were obtained for 164 cases and 20,313 controls. Increased risks of idiopathic sudden deafness were observed among participants who consumed two or more units of alcohol per day (OR=1.90, 95% CI=1.12-3.21), and among participants who slept less than seven hours per night (OR=1.61, 95% CI=1.09-2.37). The direct association with alcohol intake was particularly strong for the participants with profound hearing loss. There was little evidence of an association with smoking. This study suggests that alcohol intake and short sleep duration might be risk factors for idiopathic sudden deafness.

Aims Thrombin is an important factor in the pathogenesis of thrombotic diseases. To clarify whether smoking has an effect in platelet-dependent thrombogenesis, we studied the acute effects of smoking on platelet-dependent thrombin level in smokers. Methods and Results Subjects consisted of ten smokers and nine non-smokers. Platelet-dependent thrombin level measured after overnight fasting was greater in smokers than in non-smokers (smokers vs non-smokers, 12147 vs 56 5m IU .m l 1 , P

SummaryPlasmin inhibitor (PI) is a major physiological inhibitor of plasminmediated fibrinolysis; hence, its deficiency results in a severe haemorrhagic diathesis. We analyzed the PI gene of a French boy apparently homozygous for PI deficiency and his heterozygous parents. Both alleles of the homozygous patient had a novel G to A transition at the consensus splicing donor site in the intron 2 of the PI gene. In an expression assay using the heterologous cells transfected with the mutant PI expression vector, 3 types of aberrant transcripts using a cryptic splicing donor site within the intron 2 were detected. All of these mRNAs had a stop codon upstream of the cryptic splicing site and encode only 25 amino acids, comprising the first 21 amino acids of the signal peptide (27 amino acids) plus 4 new amino acids. This mutant was designated as PI-Paris-Trousseau.

The purpose of this study was to angioscopically observe the process of thrombolysis after intracoronary administration of thrombolytic agents and to investigate the effects of these agents on coagulation/fibrinolysis systems in dogs. The coronary endothelium was removed and thrombus formation was confirmed by angioscopy. In the tissue plas minogen activator (tPA) group (n=8), complete thrombolysis occurred in all animals, but thrombolysis was incomplete in the urokinase (UK) group (n=6). The plasma level of plasmin α2-plasmin inhibitor complex peaked at 15 minutes after treatment in both the tPA and UK groups. Plasma thrombin-antithrombin III (TAT) complex decreased tran siently at 15 minutes after tPA administration but increased at 30 and 60 minutes (P

This study investigated the clinical significance of hypoxemia without apparent clinical congestive heart failure in patients with acute myocardial infarction (AMI). Sixty-two patients with AMI of the Killip group I and Forrester subset I state were stratified into a hypoxemia group and a normoxemia group. The increase in the neutrophil count and the severity of the coronary artery disease as graded by Gensini’s score were significantly higher in the hypoxemic group. The cardiac index was lower in hypoxemic than normoxemic patients. Myocardial scintigraphy revealed no acute difference in defect scores (DS) or left ventricular ejection fraction (LVEF) between the two groups, but DS was significantly higher (p < 0.01) and LVEF was lower (p < 0.01) in the hypoxemic group 2 years after infarction. Patients with hypoxemia have a more severe angiographic coronary pathology than normoxemic patients, and latent cardiac hypofunction occurs.

A heterozygote for congenital deficiency of plasma plasmin inhibitor had hemorrhagic episodes repeatedly after the age of 79. Before the age of 79, he had not exhibited any hemorrhgic tendency and did not have abnormal bleeding even after surgical operations. Although heterozygotes for congenital deficiency of this inhibitor usually have no or only a mild hemorrhagic tendency, this case suggests that they may exhibit severe hemorrhagic tendency when they reach advanced ages because of age-related vascular changes producing hemostatic imbalance.

Objectives. This study sought to assess the usefulness of platelet-dependent thrombin generation as an index of coagulability in diabetes and to determine the effect of glycemic control on coagulability in diabetes.Background. It is important to investigate the interaction of platelets and the coagulation factors to clarify the processes of the coagulation system in detail.Methods. Platelet-rich plasma (150 × 109/liter), 0.5 ml, was prepared, and 40 mmol/liter of calcium chloride was added to initiate clotting, S-2238 was added to each sample in a microtiter plate every 10 min, and the absorbance of the released color product at 2 min was measured spectrophotometrically at a wavelength of 405 nm using a microtiter plate reader as thrombin generation. We measured the platelet-dependent thrombin generation in patients with non-insulin-dependent diabetes mellitus grouped according to glycemic control.Results. Platelet-dependent thrombin generation at 30 min after calcium chloride addition was significantly higher in 23 patients with poorly glycemic-controlled non-insulin-dependent diabetes mellitus without complications, such as diabetic retinopathy, nephropathy and neuropathy (hemoglobin [Hb] A1c⩾9.0%) than in 46 healthy normal subjects (448 ± 75 vs. 165 ± 28 mU/min, p < 0.001). Thrombin generation in 31 well controlled diabetic patients without complications (Hb A1c

The expressions of thrombomodulin (TM) and tissue factor (TF) by all- trans retinoic acid (ATRA) were studied in human leukemic cell lines including NB4 (acute promyelocytic leukemia) and U937 (monoblastic leukemia). ATRA remarkably upregulated TM antigen expression in cell lysates as well as TM cofactor activity on the cell surfaces of NB4. The level of TM mRNA in NB4 cells was increased by ATRA. Inherently procoagulant NB4 cells contained markedly higher content of TF, which was efficiently reduced by ATRA. Modest increase of TM and decrease of TF were observed when NB4 cells were treated with dibutyryl cyclic adenosine monophosphate (dbcAMP). On the other hand, both ATRA and dbcAMP showed dramatic increase of TM antigen level and modest decrease of TF antigen in U937 cells. These results suggest that ATRA regulates expressions of TM and TF antigens and activity in NB4 and U937 cell lines, and provide evidence for a potential efficiency of ATRA as a preventive and therapeutic agent for disseminated intravascular coagulation in promyelocytic and monocytic leukemia.

A lymphoma cell line with a variant type of translocation, t(2;3)(p11;q27), was established from a patient who had received liver transplantation. To elucidate the molecular mechanism of the t(2;3)(p11;q27) chromosomal translocation, we compared the structures of both derivative (der) chromosomal breakpoints with those of their germline predecessors. We noted that the BCL6 gene on chromosome 3 was juxtaposed with the immunoglobulin kappa light chain (Ig kappa) gene on chromosome 2 in a head-to-head configuration. The breakpoint of the BCL6 gene was within a previously reported breakpoint cluster region. The breakpoint on chromosome 2 was within the intron between the leader (L) and variable (V) sequences of one of the V kappa genes, which was fused to the J kappa 3 (J = joining) segment. At chromosomal junctures, a direct repeat duplication of chromosome 3 sequences and a deletion of chromosome 2 sequences were discovered. These results are consistent with a translocation model with illegitimate pairing of staggered double-stranded DNA breaks at 3q27 and 2p11, repair, and ligation to generate der(3) and der(2) chromosomes.

To investigate the clinical significance of determination of plasma tissue factor (TF) antigen, we have developed a highly sensitive enzyme-linked immunosorbent assay (ELISA) for plasma TF, using two different monoclonal antibodies against TF apoprotein, 6B4 (catching antibody) and 5G9 (detecting antibody), and tetramethyl benzidine/H2O2 as substrates. Titration curves of recombinant human TF in buffer containing Triton X-100 were linear within the range from 50 to 2000 pg/ml. The total assay time was 3 h. Ultracentrifugation and immunoblot analysis indicated that human plasma and urine contained 50,000 g sedimentable and non-sedimentable forms of TF, both of which were detected by our ELISA method. Plasma and urine concentrations of TF in healthy subjects and patients with various diseases were measured by the ELISA method. In healthy subjects, plasma and urinary TF levels were found to be 149 +/- 72 pg/ml (n = 30) and 175 +/- 60 pg TF/urine creatinine mg (n = 95), respectively. TF was increased in plasma of patients with disseminated intravascular coagulation (DIC), thrombotic thrombocytopenic purpura, vasculitis associated with collagen diseases, diabetic microangiopathy and chronic renal failure receiving haemodialysis, but not in the plasma of endotoxaemic patients without DIC. The plasma TF/serum creatinine ratio did not show a positive correlation. Measurement of TF antigen in plasma may be useful for evaluating the endothelial damage and cell destruction in TF-containing tissues.

Reciprocal exchanges between chromosomal region 3q27 and three loci of the Ig genes have been reported in cases of B-cell type non-Hodgkin's lymphoma. We have cloned a region containing a breakpoint junction of 3q27 from a cell line established from a patient with Burkitt's lymphoma carrying t(3;22)(q27;q11). The region cloned was shown to contain an Ig lambda light chain gene fused to a gene on chromosome 3q27. This finding was subsequently confirmed by fluorescence in situ hybridization. Extra nucleotides were present at the joining site. The heptamer-like and nonamer-like sequences separated by an intervening 24 bp were present in the region corresponding to the breakpoint of 3q27, suggesting that a misrecombination in Ig gene rearrangement may be involved in the translocation. Southern blot analysis with a 3q27- specific probe showed rearrangements in three additional patients with B-cell malignancies with the t(3;14)(q27;q32). The breakpoints of all four cases clustered within a limited 3-kb region on chromosome 3q27. The region of 3q27 involved in the translocation was designated as the BCL5 locus. The transcripts from the BCL5 locus were detected in normal tissues and hematopoietic cell lines, and the increased expression of transcript of aberrant size was detected in the established cell line carrying t(3;22). These observations suggest that a gene located at 3q27 is involved in the translocation and that its deregulation plays a role in the malignant transformation of B cells.

Chromosomal translocations involving band 3q27 are the recently described nonrandom cytogenetic abnormalities in B-cell malignancies. We have previously cloned the breakpoint region of 3q27, designated as the BCL5 locus, from the B-cell line carrying the t(3;22). The cDNA for the BCL5 gene was cloned from the human liver cDNA library. The nucleotide sequencing analysis showed that the BCL5 gene encodes a potential transcription factor containing six repeats of the Cys2-His2 zinc-finger motif resembling the Drosophila segmentation gene Kruppel. The calculated molecular weight was 78.8 kD, which was supported by an in vitro transcription and translation experiment. A part of the sequence was essentially identical to that of a genomic fragment, ZNF51, previously reported to be located at 3qter. The translocation occurred in the 5′ region of the BCL5 gene, and the protein-coding exons were fused to the Ig-lambda gene in a head-to-head configuration in the cell line carrying t(3;22). The BCL5 cDNA probe detected a major transcript of 3.8 kb in Burkitt's lymphoma cell lines and an aberrant transcript in the t(3;22) cell line, whereas no transcript was detected in myeloid, monocytoid, erythroid, T-lymphoid, and Epstein-Barr virus- immortalized B-lymphoblastoid cell lines.

Thrombomodulin, a thrombin receptor on the vascular endothelial cell surface, is released into circulating blood. The plasma concentrations in patients with collagen vascular diseases, including systemic lupus erythematosus (SLE), rheumatoid arthritis, juvenile rheumatoid arthritis, Sjogren’s syndrome, systemic sclerosis, polymyositis and/or dermatomyositis (PM/DM), Behcet’s disease, and Wegener’s granulomatosis, were measured and compared with those of healthy persons. The mean plasma thrombomodulin concentrations in patients with juvenile rheumatoid arthritis, systemic sclerosis, PM/DM, Wegener’s granulomatosis, and active states of SLE, rheumatoid arthritis, and Bechet’s disease were significantly higher than those in the control group. Patients with Wegener’s granulomatosis showed the highest mean value. The mean values in patients with inactive states of the diseases and Sjogren’s syndrome were not significantly different from the values in the control group. In cases of juvenile rheumatoid arthritis, systemic sclerosis, PM/DM, and Wegener’s granulomatosis, patients with active interstitial pneumonitis or extensive pulmonary lesions frequently showed higher values of plasma thrombomodulin than those without overt pulmonary involvement. Elevated plasma thrombomodulin values were decreased along with amelioration of the diseases by treatment. These results may indicate that plasma thrombomodulin measurement may be helpful for evaluating vascular injury in patients with collagen diseases.

The c-kit proto-oncogene encodes a tyrosine kinase receptor for stem cell factor and plays a critical role in the growth and differentiation of various types of cells including hematopoietic stem cells. To investigate the mechanisms of its transcriptional regulation, we isolated the 5' flanking region of the human c-kit gene and characterized its promoter activity in hematopoietic cells. Nucleotide sequence analysis revealed that the 1.2 kb 5' flanking region lacked a typical "TATA box," but had a relatively high G + C content and four potential Sp1-binding sites. Putative binding sites for AP-2, basic helix-loop-helix proteins, Ets-domain proteins, Myb and GATA-1 were also found. Primer extension and S1 nuclease protection analyses of hematopoietic cells indicated that the major transcription start sites are 62 bp and 58 bp upstream of the translation start site. Essentially the same start sites were detected in non-hematopoietic cells such as small cell lung carcinoma and glioblastoma: this single promoter in c-kit is different from the multiple promoter system of c-fms, a c-kit-related gene, in which at least two promoters are differently used in hematopoietic and non-hematopoietic cells. An analysis of the c-kit 5' flanking region using the bacterial chloramphenicol acetyltransferase gene (CAT assay) in human erythroleukemia HEL cells, which express the endogenous c-kit mRNA at high levels, showed that a region from -180 to -22 is important for the expression of the c-kit gene. In addition, a negative regulatory element(s) is suggested to be involved in the regulation of the c-kit gene expression in mammals.

Publisher Summary This chapter describes the human α2-plasmin inhibitor. α2 PI molecule has three functional sites: the reactive site, plasminogen-binding site, and cross-linking site. The plasminogen-binding site, located within 20 amino acid residues of the carboxyl-terminal end, binds to the lysine-binding sites of plasminogen, to which fibrin is also bound. Hence, α2PI competitively inhibits the binding of plasminogen to fibrin thus retarding the initiation of the fibrinolytic process because plasminogen binding to fibrin plays an important role in the initiation of fibrinolysis. The high affinity of the plasminogen-binding site for plasmin accelerates the complex formation of plasmin and α2PI, resulting in a rapid inhibition of plasmin. The cross-linking site is located at the second amino acid residue, glutamine, from the amino terminus. The cross-linking of α2PI to fibrin plays a significant role in the inhibition of fibrinolysis because it stabilizes the fibrin clot against fibrinolysis. α2PI can be purified directly from human plasma by immunoaffinity chromatography using a monoclonal antibody.

Thrombomodulin(TM), is an endothelial cell surface glycoprotein that plays an important role in thromboresistance especially in microvasuculature. It forms a one to one complex with thrombin to inactivate its procoagulant property. This complex catalyzes activation of protein C and activated protein C inactivates coagulant factors Va and VIIIa in the presence of protein S. Thus, TM converts procoagulant thrombin to an anticoagulant and plays a central role in the protein C anticoagulant pathway. The physiological importance of the TM-protein C system in the prevention of thrombus formation is supported by various observations including thrombotic tendencies in patients with congenital deficiency of protein C and effective prevention of experimental thrombosis by intravenous administration of purified TM. Previous studies have shown that TM on endothelial cells is down-regulated by endotoxin, TNF or IL-1β. This loss of anticoagulant potential together with induction of tissue factor in blood coagulation by these agents is thought to be related to the hypercoagulative state of sepsis, chronic inflamation and cancer. Therefore, states of expression of TM on surfaces of endothelial cells are important in regulation of intravascular coagulation.

To investigate leukemogenesis of acute promyelocytic leukemia (APL), we studied the involvements of retinoic acid receptor alpha (RAR alpha) and myl genes, and also the frequency of N-RAS, K-RAS, H-RAS, and FMS point mutations in sixteen patients with APL. By Southern blot analysis, the rearrangements of RAR alpha gene were detected in 13 patients (81.2%), and myl gene in 14 (87.5%). Either RAR alpha or myl gene rearrangements were found in all patients including one with normal karyotype. Breakpoints of both genes were clustered. By direct sequencing, no point mutations were found at codons 12, 13, and 61 of N-, K-, and H-RAS genes, and at codons 301 and 969 of FMS gene. These data indicate that myl-RAR alpha translocation occurs frequently in APL, whereas RAS and FMS mutations are rare in APL. It may be suggested that leukemogenesis of APL is different from other subtypes of acute myelogenous leukemia, and multistep leukemogenesis may not be a prevalent feature in APL.

We characterized a mutant protein C gene from an individual with no detectable protein C antigen in blood plasma. Southern blot hybridization analysis with human protein C cDNA demonstrated neither gross deletion nor rearrangement of the gene. Sequencing all the exons and exon-intron boundaries of the gene except the 3′ noncoding region showed two mutant alleles. The one, derived from the mother, represents a deletion of 5 nucleotides (nt) (CCCGC) in the end of exon VI (mutation I), predicted to result in the generation of a new stop codon due to a reading frameshift and the premature termination of translation. The other, derived from the father, represents a point mutation (G to A) in exon IX (mutation II), resulting in an amino acid substitution, Gly-376(GGC) to Asp(GAC), in the catalytic domain of the protein. Allele-specific oligonucleotide probe hybridization confirmed the presence of the two mutations. Mutation I would result in a truncated polypeptide of 169 amino acid residues that lacks the heavy chain. Mutation II gives rise to an alteration of a highly conserved amino acid, Gly-376. These data indicate that this patient is a compound heterozygote of the two mutant alleles, each one inherited from each parent. Transient expression assays using COS-7 cells transfected with mutated protein C expression vectors suggested that each of the two mutations leads to the protein C deficiency by causing an impairment of secretion of the respective mutant proteins.

ala-plasmin inhibitor (cLzPI) has been purified from human plasma by conventional multistep methods (1, 2). Although the methods allow the recovery of high quality of aaP1, low overall yield and prolonged time required for purification make the methods unsuitable for routine use. In this communication, we describe a newly developed immunoaffinity purification method for claPI, which is simple, efficient and easily performed.

Thrombomodulin (TM) is a cell surface glycoprotein that plays an important role in inhibition of intravascular coagulation as a cofactor for the thrombin-catalyzed activation of protein C.1 It contributes to thromboresistant properties of endothelium. We reported that TM-fragments increased in plasma of patients with various thrombotic disorders such as DIC, ARDS and pulmonary thromboembolism.2 However, it is not clear whether the increase of TM-fragments in plasma is caused simply by vascular damages or reflects an increase of expression of TM in endothelial cells. In this paper, we summarize our studies on the regulatory mechanisms of TM expression in vitro assuming its role in pathogenesis of hypercoagulable states.

A novel human leukemic cell line, TMD2, which proliferates dependently on interleukin-3 (IL-3), was established from the peripheral blood (PB) of a patient with chronic lymphocytic leukemia (CLL) in the acute phase. After 8 years of the chronic phase of CLL, lymphoblastoid cells appeared and became dominant in the PB. After repeated subcultures of the patient's PB cells in the acute phase, lymphoblastoid cells have proliferated actively in the presence of recombinant human IL-3, and the TMD2 cell line has been established. The lymphoblastoid cells in acute phase and TMD2 cells proliferated dependently on IL-3, whereas growth of the small lymphocytes in chronic phase was not supported by IL-3. Other ILs (IL-1, 2, and 4 through 6) or CSF did not support the growth or survival of TMD2 cells. The existence of high-affinity receptors for IL-3 was shown on TMD2 cells (binding sites 88 per cell, Kd = 76.9 pmol/L). DNA extracted from the small lymphocytes in the chronic phase, the lymphoblastoid cells in the acute phase, and TMD2 cells showed the same rearrangement pattern of the immunoglobulin heavy- chain gene. Therefore, these cells were considered to have originated from the same clone. These results imply that a genetic event that caused the responsiveness to IL-3 in the cell of the chronic phase caused the acute transformation of CLL in this patient. We consider that TMD2 cell line is valuable as a model of cells of CLL in the acute phase and as a tool for studying the signal transduction system of IL-3.

The effect of recombinant human interleukin-4 (IL-4) on a granulocyte colony-stimulating factor (G-CSF)-dependent human myeloblastic leukemic cell line, OCI-AML1a, was investigated. IL-4 suppressed the clonogenic cell growth in methylcellulose culture, inhibited the uptake of 3H thymidine in a dose-dependent manner at 5 to 100 U/mL, and consequently suppressed the growth of clonogenic cells in short- and long-term suspension cultures. In addition, IL-4 markedly increased the number of adherent cells. These adherent cells were alpha-naphthyl-butyrate (alpha-NB) esterase-positive and showed macrophage-like appearance, increased expression of CD14, CD11b, CD23, and Ia, and significantly decreased clonogenicity. On the other hand, nonadherent cells growing in suspension showed only slight increase in proportion of alpha-NB esterase-positive or monocyte/macrophage-like cells and increased CD23 expression by an addition of IL-4. The clonogenicity of the nonadherent cells was not significantly influenced by IL-4. By addition of the media conditioned by OCI-AML1a cells in the presence of IL-4, the clonogenic cells growth of OCIAML1a cells was suppressed and adherent cells were markedly increased. The suppressive and differentiative effects on OCI/AML1a cells of the conditioned media and IL-4 itself were almost completely abolished by anti-IL-4 antibody. Furthermore, the neutralizing antibodies against transforming growth factor-beta 2 (TGF-beta 2), tumor necrosis factor-alpha (TNF-alpha), or IL-6 did not influence the effect of recombinant IL-4. Taken together, IL-4 was shown to suppress the growth and induce differentiation toward adherent macrophage-like cells of the G-CSF-dependent myeloblastic cell line. The effect of IL-4 may be direct, and not secondary via inducing production of other cytokines such as TGF-beta, TNF-alpha, or IL-6 by leukemic cells.

Let M be a closed manifold and f: M → M be a Cr diffeomor-phism. Let p ∈ M be a hyperbolic fixed point of f. Then the stable and unstable sets of p are denoted respectively by $$ \begin{array}{*{20}{c}} {{W^S}\left( p \right) = \left\{ {x\left| {\mathop {\lim }\limits_{n \to \infty } } \right.d\left( {{f^n}\left( x \right),p} \right) = 0} \right\}} \\ {{W^u}\left( p \right) = \left\{ {x\mathop {\left| {\lim } \right.}\limits_{n \to \infty } d\left( {{f^{ - n}}\left( x \right),p} \right) = 0} \right\}} \\ \end{array} $$ which are Cr injectively immersed submanifolds of M. The points of intersection of the closure of Ws(p) with Wu(p) or of the closure of Wu(p) with Ws(p), different from p, is called almost homoclinic points to p.

The effects of recombinant human interleukin-6 (rh IL-6), which has homology with rh granulocyte colony-stimulating factor (rh G-CSF) at the amino acid sequence level, and rh G-CSF on normal human bone marrow cells, fresh leukemic blast progenitors from 16 acute myeloblastic leukemia (AML) patients, and G-CSF-dependent human AML cell line (OCI/AML 1a) were investigated. rh G-CSF stimulated the proliferation of leukemic blast progenitors from 13 out of 16 AML patients tested. rh IL-6 stimulated the proliferation of blasts from eight AML patients and enhanced the G-CSF-dependent proliferation of the fresh AML blasts from two out of eight patients tested. On the other hand, rh IL-6 suppressed the blast colony formation from two AML patients and OCI/AML 1a cells and also reduced the G-CSF-dependent proliferation of the blast progenitors from one of the two patients and the cell line, rh IL-6 had no effect on the colony formation of normal granulocyte-macrophage colony-forming units (CFU-GM) with or without rh G-CSF. Differentiation-induction by rh IL-6 was not observed in the fresh AML blasts but was observed in OCI/AML 1a. The effect of IL-6 on the blast colony formation and G-CSF-dependent blast cell growth was complicated and heterogenous among the AML cases; IL-6 stimulated blast colony formation in some cases and suppressed it in others. The heterogeneity of the response was supposed to be derived from the heterogeneity of the characteristics of AML cells. Although G-CSF simply stimulated the blast colony formation, IL-6 had a bimodulatory effect on the proliferation of leukemic blast progenitors from AML patients. IL-6 might be involved in the regulation of the proliferation of AML cells in vivo as well as in vitro.

Defibrotide is known to enhance prostacyclin (PGI2) release from the vascular endothelium. We investigated the vasoactive effects of defibrotide in isolated rat hearts perfused at constant flow subjected to ischaemia and reperfusion. Defibrotide at 10 or 100 micrograms/ml did not exert any direct vasoactive effect on normal rats hearts. However, ischaemia and reperfusion resulted in an impaired vasodilation to acetylcholine, an endothelium-dependent vasodilator. In contrast, the vasodilator response to the endothelium-independent dilator, nitroglycerin, was unaffected. Defibrotide, at 10 or 100 micrograms/ml, markedly restored the vasodilation to acetylcholine 10 nmol/l to 1 mumol/l (P less than 0.01) without influencing the vasodilator response to nitroglycerin (2 to 200 micrograms/l). Haemoglobin (150 nmol/l) inhibited the dilation to acetylcholine in response to defibrotide. However, no evidence of PGI2 release was observed with acetylcholine-induced vasodilation in the presence or absence of defibrotide. Additionally, 10-100 micrograms/ml of defibrotide did not significantly decrease superoxide radicals generated by a xanthine-xanthine oxidase synthetic system under conditions in which superoxide dismutase was effective. Thus, defibrotide appears to exert an endothelium-protective effect preserving endothelium-derived relaxing factor (EDRF) without directly scavenging free signals.

SummaryAlthough protein C (PC) and activated protein C (APC) have been postulated to be useful for treating patients with thrombosis, their critical effect remains to be studied in human subjects. To examine whether purified PC or APC are useful for treating patients with thrombosis without showing any adverse effect, wc studied effects on coagulation and fibrinolysis in normal human subjects.When highly purified human PC was administered intravenously to healthy subjects, plasma levels of immunoreactive PC decreased with a half-life of 10.9 h. Intravenously administered APC decreased with a half-life of 23 min as measured by prolongation of activated partial thromboplastin time (APTT). However, 1.7 h was obtained for the plasma half-life of APC when it was measured immunologically. These findings suggested that a significant fraction of the administered APC was rapidly inhibited by plasma inhibitor. Upon administration of APC, APTT was prolonged and plasma levels of clotting factor VIII (FVIII) decreased transiently as measured by clotting assay. However, when determined by a chromogenic assay method in which 120-fold diluted plasma samples were used, plasma levels of FVIII remained unchanged. Plasma levels of F-V did not decrease after APC administration. These findings suggested that prolongation of APTT and apparent decrease in plasma F-VIII clotting activity might be due to the in vitro-effect of APC present in plasma samples used. Diurnal fluctuation of plasminogen activator inhibitor in normal subjects was not affected by administration of APC.Thus, PC or APC seems to function selectively at the site of thrombin-formation without lowering plasma levels of coagulation factors.

α 2-Plasmin inhibitor (α 2PI), also called α 2-antiplasmin, is a plasma glycoprotein which rapidly inactivates plasmin proteolytic activity (1). Its molecular weight, deduced from the cDNA sequence and the carbohydrate content (14%), is ≅ 58 KD (2) whereas the molecular weight estimated by SDS-gel electrophoresis was 67 KD (1). The cause of the discrepancy is not known. Its concentration in human plasma was estimated to be 6.9 ± 0.6 mg/100 ml (3), which is estimated to be ≅ 1.2 μM on the assumption of MW 58KD. α 2PI is a serine proteinase inhibitor (serpin) which is able to inhibit several different “serine” proteinases, but is mainly playing a role as a primary inhibitor of plasmin-mediated fibrinolysis (4). Its congenital deficiency results in a lifelong severe hemorrhagic tendency due to premature degradation of hemostatic plugs by physiologically occurring fibrinolytic process (4–7).

Endothelin, a newly discovered vasoconstrictor peptide, when added to isolated cat papillary muscles, induced a direct positive inotropic effect that was slow in onset but of long duration. The magnitude of the developed force was concentration dependent. Endothelin exerted a marked concentration-dependent vasoconstriction in isolated cat carotid arteries and rabbit aortic rings. In both the carotid arteries and the aortic rings, endothelin induced a similar vasoconstrictor effect in the presence or absence of an intact endothelium. Addition of propyl gallate, a 5-lipoxygenase inhibitor, ibuprofen, a cyclooxygenase inhibitor, or SKF-525A, a cytochrome P450 inhibitor, at 2 microM did not significantly attenuate the ability of endothelin to vasoconstrict aortic rings in the presence or absence of an intact endothelium. These results demonstrate that the vasoconstrictor activity of endothelin operates independently of all three pathways of arachidonic acid metabolism (i.e., lipoxygenase, cyclooxygenase, or cytochrome P450 pathways) and is not dependent upon other endothelium-derived mediators (e.g., endothelium-derived relaxing factor, or eicosanoids) in these preparations. Moreover, endothelin exerts a direct positive inotropic effect in isolated cat ventricular myocardial tissue.

In a previous work, authors demonstrated a new carbon corrosion mode caused by rapid voltage rises in PEFC cathode. In this study detailed investigations for the carbon corrosion are conducted with full-scale cells. Test results suggest that higher potential sweep rate and larger potential gain which relates to rapid voltage raises during start-up could accelerate carbon corrosion significantly, without local or global fuel starvation. Impacts of carbon corrosion on material degradation and cell performance decay are also evaluated. The results indicate that voltage cycles between start-up and shutdown, which advances carbon corrosion on the cathode, accelerates cell performance decay with electrochemical surface area reduction and mass transport loss on the cathode. The voltage cycle testing focusing on carbon corrosion is expected to be one of the most promising test protocols for accelerated durability testing.

Background: Thrombomodulin is a thrombin receptor on the endothelial cell surface that plays an important role in the regulation of intravascular coagulation. Recombinant human soluble thrombomodulin (ART-123) is composed of the active, extracellular domain of thrombomodulin. ART-123 has been shown to have a wider safety margin than other anticoagulants and to have a favorable antithrombotic profile with less bleeding in animal and in vitro experiments. ART-123 is a promising therapeutic natural anticoagulant that is comparable to antithrombin, tissue factor pathway inhibitor and activated protein C. Objective: We conducted a multicenter, double-blind, randomized parallel-group trial to compare the efficacy and safety of ART-123 to those of heparin for the treatment of disseminated intravascular coagulation (DIC) associated with infection or hematologic malignancy. Methods: DIC was diagnosed according to the diagnostic criteria established by the Japanese Ministry of Health and Welfare. DIC patients were assigned to receive ART-123 (0.06 mg/kg for 30 min, once a day) or heparin sodium (8 units/kg/h for 24 h) for six days using a double dummy method. The primary efficacy endpoint was DIC resolution rate (rate of recovery from DIC). The secondary endpoints included clinical course of bleeding symptoms, and mortality at 28 days. Results: 234 DIC patients were randomized (117 in each arm). DIC resolution rate was 66.1% for the ART-123 group and 49.9% for the heparin group (difference 16.2%; 95% CI 3.3–29.1), thus demonstrating that ART-123 is significantly superior to heparin for the improvement of DIC. Patients in the ART-123 group also showed more marked improvement in clinical course of bleeding symptoms (p=0.0271) and the disappearance rate of bleeding symptoms in the ART-123 group was higher than that in the heparin group (35.2% vs 20.9%; difference 14.3%). The incidence of bleeding-related adverse events up to 7 days after the start of administration was lower in the ART-123 group than in the heparin group (43.1% vs 56.5%; difference; −13.4%; p=0.0487). The mortality rate at 28 days in the ART-123 group was 22.0% vs 25.5% in the heparin group (difference −3.4%; p=0.5396). Although no significant differences were seen, 28-day mortality for the ART-123 group was slightly lower. The mortality rate of patients with DIC secondary to infection in the ART-123 and heparin groups was 28.0% (14/50) and 34.6% (18/52), respectively, indicating a 6.6% lower mortality in the ART-123 group. Conclusion: When compared with low-dose heparin, ART-123 more significantly improves DIC and alleviates bleeding symptoms in DIC patients. The incidence of bleeding-related adverse events is significantly lower with ART-123 than with heparin. Because of its safety and efficacy, ART-123 appears to be a first-line agent in the management of DIC.

Carbon corrosion caused by rapid potential changes is presented. Test results with full-scale single cells suggest rapid voltage changes during start-up and shutdown can accelerate carbon corrosion significantly, without local or global fuel starvation. Impacts of carbon corrosion on material degradation and cell performance decay with various operation modes (e.g. voltage cycles, OCP hold and constant load) are also evaluated. The results indicate voltage cycles by start-up and shutdown, which advances carbon corrosion on cathode, accelerates cell performance decay with electrochemical surface area reduction and mass transport loss on cathode. The voltage cycle testing focusing on carbon corrosion is expected to be one of the promising test protocols for accelerated durability testing.

We previously assigned the human α2-plasmin inhibitor gene (PLI) to 18p11.1→q11.2 by isotopic in situ hybridization. However, results obtained subsequently via Southern blot analysis of somatic cell hybrids and fluorescence in situ hybridization studies indicate that the gene is located at 17p13. Accordingly, we reassign PLI to 17p13.

The therapeutic effect of highly purified protein C (PC) or activated protein C (APC) on three patients with disseminated intravascular coagulation (DIC) was tested. In two out of three cases, although therapeutic dose of heparin was not effective, PC or APC administration significantly improved the hypercoagulable state. In one case with acute leukemia who developed DIC with the severe gastrointestinal bleeding, APC corrected the hypercoagulable state without aggravating bleeding. These findings suggested that PC and/or APC might be effective in correcting hypercoagulable state without any adverse effect.

The purpose of this paper is to prove the fact stated by the title. After Ornstein and Friedman [5], [1], several authors studied the Bernoulli properties of various transformations. Especially Katznelson [3] proved that every ergodic automorphism of a finite-dimensional torus is a Bernoulli transformation extending the results by Sinai-Ornstein-Friedman [9], [1]. The present result is on the way towards the conjecture that every ergodic automorphism of a compact metrizable abelian group is a Bernoulli transformation. Let X be a compact metrizable group and /^ be its normalized Haar measure. Then (X, ft) is a Lebesgue space (cf. [10]). Let a be an (group) automorphism of X, then cr is an invertible measure-preserving transformation of (X, fji). Our problem is concerned with measure-theoretic properties of a. We call a a Bernoulli transformation if there exists a measurable partition f of X such that {o-^}_00?/<00 are independent and V o-f=£ the partition of X into individual points. We assume X—T°° an infinite-dimensional torus i.e. X is a compact metrizable abelian group which is connected, locally connected and infinite-dimensional. Further we assume naturally that the automorphism a is ergodic i.e. any cr-invariant measurable set has measure 0 or 1. Our result is the following

Transparent halopolymer film such as poly(vinyl chloride) containing leuco dye formed color image by UV irradiation. Combinations of six kinds of polymers and five kinds of leuco dyes were investigated as photo coloring systems. A stable coloration of the leuco dye can be obtained if halopolymer is used as a matrix film. The transparent film of poly(vinyl chloride) (PVC) which contained leuco auramine benzotriazole (1-ABT) changed to greenish blue by UV irradiation for about 10sec. Photo-sensitivity of 1-ABT depended on the kind of halopolymer films. Poly(vinyl chroride-co-acrylonitrile) gave high photosensitivity for color formation of 1-ABT. Color formation of 1-ABT in poly(2-chloroacrylonitrile)(PCAN) by UV light was very fast, and was observed during casting the film under room light. Therefore only photobleaching was observed in film state. Color formation of 1- ABT with PCAN was also investigated in acetonitrile solution and in dimethylformamide solution. Color formation in dimethylformamide was faster than in acetonitrile. Photochemical reaction of fluoran lactone (FL) in PCAN film gave interesting result. Slightly greenish film of the FL-PCAN system colored quickly to red by irradiation, followed by changing to black by continuous irradiation. The UV and IR spectra of the PCAN film containing FL under UV light suggested formation of poly(cyanoacetyle).

A patient who suffered a recurring thrombosis over the last 15 yr has been investigated. The only abnormality found in this patient was a significantly depressed level of plasminogen activity in plasma. In spite of the depressed plasminogen activity, the patient was found to have a normal level of plasminogen antigen concentration. It was calculated that the activity per milligram of plasminogen of the patient was approximately one-half the values of normal subjects. The same discrepancy between biological activity and antigen concentration was found in the other members of the kindred. A niece was found to have practically no plasminogen activity but possessed a normal concentration of plasminogen antigen. Both her parents were found to have approximately half the normal plasminogen activity and normal antigen levels. These studies suggested that the molecular abnormality was inherited as an autosomal characteristic, and the family members who had half the normal levels of activity with normal plasminogen antigen were heterozygotes whereas the one with practically no plasminogen activity was homozygote. Subsequent studies showed that the pattern of gel electrofocusing of purified plasminogen of the heterozygotes consisted of 10 normal bands and 10 additional abnormal bands, each of which had a slightly higher isoelectric point than each corresponding normal component. This indicates that plasminogen of the heterozygote is a mixture of normal and abnormal molecules in an approximately equal amount, which was substantiated by active site titration of purified plasminogen preparations obtained from the propositus and a normal individual. The gel electrofocusing pattern of the homozygote consisted of abnormal bands only. The defect is a hereditary abnormality of plasminogen.

In the first part of this study, oxygen derived radicals (O2-radical) were indirectly demonstrated as causative agents for ischemia/reperfusion injury by showing myocardial protection of radical scavengers. In 19 open chest dogs, the left anterior descending coronary artery was occluded for 90 min and subsequently reperfused for 60 min. Group A received SOD (15000 U/kg), catalase (5 mg/kg) and 20% mannitol (18 ml/kg) via the left atrium starting 15 min pre-occlusion and ending 15 min after reperfusion. Group B received infusion of saline and served as control. The severity of myocardial injury was evaluated by epicardial ECG, hemodynamics, echo cardiographical asynergy area and left ventricular wall thickness, histopathological findings and magnitude of necrotic area. Group A demonstrated less injury than group B in most of the parameters. In particular, ratios of necrotic to perfused areas determined by dual fluorescence methods were significantly limited in group A (34.1 +/- 12.0 vs 66.0 +/- 11.3%), indirectly verifying that the O2- radicals play an important role in the genesis of ischema/reperfusion injury. In the second part of the study, the direct measurement of radicals in freeze-clamped myocardium was reported using electron spin resonance (ESR) spectroscopy. With this method, it was not possible to demonstrate the generation of O2-radicals in ischemic/reperfused myocardium. However, the data suggested that coenzyme Q10 anion radical, which exists in the normal myocardium, might provide an index of tissue injury.