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To investigate whether telmisartan (TMS), a unique angiotensin II type 1 receptor(AT1R) antagonist with selective peroxisome proliferator-activated receptor (PPAR) γ- modulating activity, inhibits plasminogen activator inhibitor 1 (PAI-1) and/or stimulates issue-type plasminogen activator (t-PA) mRNA expression in vascular endothelial cells(VECs), we cultured VECs obtained from the thoracic aorta of Long Evans Tokushima Otsuka rats by the explant method. mRNA expression was measured using comparative reverse transcription polymerase chain reaction. The binding activity of PPARγ to its coactivator (CREB-binding protein) was determined by enzyme-linked immunosorbent assay. The synthetic PPARγ agonists troglitazone (TRO) and TMS activated PPARγ with EC_50 values of 0.95 and 161μM, respectively, under cell-free conditions. PPARγ mRNA was constitutively expressed in cultured VECs, unlike PPARα and PPARβ. PAI-1 mRNA expression was significantly downregulated by TMS, and this downregulation was not abolished by the presence of the PPARγ antagonist GW9662. The expression of t-PA mRNA was not altered by TMS treatment. TRO had no effect on PAI-1 and t-PA mRNA expressions.PAI-1 mRNA expression was significantly decreased by GW9662. In conclusion, TMS inhibited PAI-1 mRNA expression independently of PPARγ activation. Intrinsic PPARγ agonists, but not extrinsic PPARγ agonists, may be involved in basal mRNA expression of PAI-1 in VECs

The aim of this study was to find the relationship between change in taste and food intake in the cancer patients underwent chemotherapy. Gustatory thresholds were measured by dropping the taste liquid on the area innervated by the chord tympani nerve in 31 patients underwent chemotherapy in two hospitals. Taste tests, measurement of food intake and questionnaires were performed four times : before the chemotherapy (Before), on the 3 rd day (3 rd) and the 7 th day (7 th) the chemotherapy, and at 2 weeks after the termination of chemotherapy (After). Forty-eight percent of the patients reported taste changes in the course of chemotherapy. The most frequent complaints were "feel lighter taste (60.0%)" and "become sensitive to sweetness (40.0%)" were followed. Taste test revealed that the percent of change in gustatory threshold for salty was significantly increased on 3 rd, whereas that for sweet was significantly decreased on After. The percent of food intake was significantly decreased on 3 rd (72.3%) and 7 th (78.3%) as compared to Before (84.2%), and was gradually recovered on After. The percent of food intake was significantly decreased in the patients complaining taste change. Furthermore, The cancer patients with higher gustatory threshold for salty tend to less the percent of food intake. In conclusion, taste change was found in the early during chemotherapy, its relation with the amount of food intake was also demonstrated in the cancer patients underwent chemotherapy. Therefore, we would like to propose that we need to assess taste change in the early during chemotherapy to support food intake in the cancer patients, for example, serving salty meal than usual in order to prevent decrease in amount of food intake during chemotherapy.

To investigate whether a feeding action of ghrelin and an antiobese action of leptin are modified by cholecystokinin (CCK), body weight, food intake, and plasma concentrations of CCK, active ghrelin, desacyl ghrelin and leptin were measured in 7 and 38 weeks old of Otsuka Long-Evans Tokushima Fatty (OLETF) rats which lack CCK1 receptor. Long-Evans Tokushima Otsuka (LETO) rats were used as normal counterpart. The mean body weight, the amount of food intake, active ghrelin and leptin were significantly increased, whereas CCK was significantly decreased in OLETF rats as compared to that in LETO rats at 7 and 38 weeks old. Multiple linear regression analysis revealed that active ghrelin and leptin were significant determinants for prediction of food intake in OLETF rats and that leptin was a significant determinant for body weight in LETO rats. Leptin resistance index (LRI) was calculated by a following equation; LRI = body weight x plasma leptin. The index was negatively correlated with concentration of CCK.Thus, the decreased plasma CCK in OLETF rats may associate with leptin resistance and increase body weight.

We investigated whether elevation of intracellular cyclic nucleotides (cAMP and cGMP) inhibit cell proliferation and migration of cultured embryonic rat aortic vascular smooth muscle (VSM) cells (A7r5). After treatment with drugs for up to 120 h, cell proliferation and migration were measured by adding WST-1 reagent and by counting migrated cells through the pores of cell culture insert (8-$ \mu $ pore size) , respectively. Cell proliferation was significantly inhibited by dibutyryl cAMP (db-cAMP) in a time- and concentration-dependent manner (0.03 to 3 mM) , whereas dibutyryl cGMP (db-cGMP, 3 mM) had no effect. Olprmone hydrochloride (OPN, 1 mM) , type 3 phosphodiesterase inhibitor, inhibited cell proliferation, and synergistically inhibited in the presence of db-cAMP (0.3 mM). Cell migration was significantly inhibited by treatment with db-cAMP (3 mM), but not by db-cGMP (3 mM). Combination of db-cAMP (0.3 to 3 mM) with OPN (1 mM) or forskolin (FSK) (0.1 mM) significantly inhibited cell migration after 72-htreatment. Plateletderived growth factor ( PDGF) -BB significantly stimulated cell proliferation and migration in serum-free culture condition. Treatment of the cells with db-cAMP, OPN or FSK significantly inhibited the stimulatory effect of PDGF-BB on proliferation and migration. Thus, elevation of cAMP inhibited serum stimulated and PDGF-BB induced cell proliferation and migration of A7r5 cells.

Many protease inhibitors in the circulating blood inhibit the activity of $ \alpha $-thrombin. Inhibition of $ \alpha $-thrombin by antithrombins is attributable to the characteristics of structural relationships between $ \alpha $-thrombin and antithrombins. The characteristic structures of $ \alpha $-thrombin include loop 60, exosites 1 and 2, hydrophobic pocket, primary specificity pocket, and active site (catalytic triad). Functions of these structures have been clarified by enzyme-substrate reaction and X-ray and NMR-crystallographic analyses. For developing antithrombine drugs that is safely and effectively used for clinical treatment of thrombotic diseases, the precise understandings of relationship between structure and function of α-thrombm are required. Although it has been evidenced that clot-bound thrombm plays more important role in pathophysiological conditions than native (free) thrombin, the structure and function of clot-bound thrombin remains to be clarified. Understanding the structure and function of clot-bound thrombm would provide a potential therapeutic strategy to develop novel antithrombin drugs, which control the activity of the clot-bound thrombin in the circulating blood. In this review paper, first of all we summarized the structure-function relationships based on the studies on native thrombin and mutant thrombin. Secondary, we summarized the mechanism for thrombin receptor activation by $ \alpha $-thrombin and thrombin receptor agonist peptide (TRAP). Finally, we demonstrated our resuits of the studies on the characteristics of clot-bound thrombin

Procoagulant activity was known to increase after thrombolytic therapy. It was thought that the enhancement of procoagulant activity was dependent upon the new generation and development of novel kinds of thrombin after thrombolytic therapy. Such thrombin was also produced by percutaneous transluminal coronary angioplasty (PTCA) and percutaneous transluminal coronary reperfusion (PTCR) , resulting in unexpected rethrombosis and vascular restenosis. That is, thrombin appeared again in the circulating blood after thrombolysis or mechanical destruction of the previously formed thrombus. Such the newly generated thrombin was termed postclotting thrombin (bound thrombin). The postclotting thrombins were divided into three classes: native, intact, and bound thrombin. It was emphasized in this review that the bound thrombin generated during the formation of fibrin or thrombus has the clotting activity like a native thrombm. The reliable control of the bound thrombin using synthetic antithrombin agents led the clinical doctors to reasonable haemostasis, tissue repair and regeneration, and prevention from atherosclerotic change of blood vessel

We have reported that habutobin acted only on rabbit fibrinogen by cleaving the Arg^16-Gly^17 bond in the A$ \alpha $ chain. We have also demonstrated that habutobin recognized threonine (Thr) at position 10 from the COOH-end of the rabbit fibrinopeptide A (FPA). Since Thr has been revealed to also exist at position 10 in chicken FPA, we postulated that habutobin may recognize chicken Thr, thereby converting it into fibrin gel. We investigated whether habutobin acted on chicken fibrinogen to form fibrin gel. Habutobin clotted chicken fibrinogen more slowly than it did rabbit fibrinogen. It clotted the chicken fibrinogen with a resultant release of three peptides (peak I, II and X peptides), whereas thrombin clotted the chicken fibrinogen with a resultant release of two peptides (peak 1 and 2 peptides). The elution times of the peak I and II peptides were identical to those of peak 1 and 2 peptides. The peak 2 fibrmopeptide was identified as the FPA of chicken fibriogen. A computer-assisted structural analysis of the rabbit FPA demonstrated that the Thr is located at the top of the N-termmal cluster, whereas the Thr in the chicken FPA exists in a linear and flexible peptide. Since the Thr on the chicken FPA seemed to fluctuate in the peptide, recognition of the Thr by habutobin might be unstable and ineffective. In conclusion, the exposure of Thr at position 10 in the FPA molecule is essential for habutobin-specific recognition of chicken fibrinogen.

Both percutaneous transluminal angioplasty (PTA) and percutaneous transluminal coronary angioplasty (PTCA) are useful for the treatment of patients with vascular stenosis. However, restenosis occurs among 30% to 40% of those treated within six months. A variety of experimental and clinical studies have been directed toward the clarification of the pathogenesis of restenosis, which have led to the development of new methods to prevent restenosis with partial success. In this paper, from the point of view of coagulation-fibrinolysis system, we summarized the pathognomotic factors that develop restenosis and the agents that prevents restenosis from the previously cited literatures. In addition, we stressed that postclotting thrombins ineluding clot-bound thrombin that could be generated in the case of mechanical medical procedures and a fibrinolytic therapy for coronary stenosis, play an important role in the development of rethrombosis and restenosis., 論文

Changes in the cytoskeletal network alter the mechanical properties of cells that are essential for functions such as locomotion and cytokinesis. The transmembrane receptors and intracellular signals have been extensively studied on cytoskeletal changes in response to extracellular stimuli. After the procedure of percutaneous transluminal coronary angioplasty (PTCA) and highfrequency rotational atherectomy (PTCR) for stenosis of blood vessels, restenosis may be observed. The restenosis were generated under the proliferation and migration of vascular smooth muscle (VSM) cells. In addition, until recently it has been expected that the physiological regulation of VSM cells was dependent on L-type Ca^ (Ca_1) channel activity and structure of the cytoskeleton. To clarify the interactions between the structure of the cytoskeleton and activity of Ca_1 channel in VSM cell, we used electrophysiological and morphological experiments concomitantly. We investigated the effect of disruption of the actin filaments and the microtubules on Ca_1 channel activity (Ca^ current) of cultured VSM cells (A7r5 cell line) using whole cell voltage clamp. The results of immunostaining using anti-$ \alpha $-actin and anti-$ \alpha $-tubulin antibodies showed that colchicines disrupted both actin filaments and microtubules, while cytochalasin D and nocodazole disrupted only actin filaments, and microtubules respectively. Ca^ current was greatly reduced in cells that were exposed to colchicines or cytochalasin D but not to nocodazole. When the actin filaments were stabilized by phalloidine, the inhibition of Ca^ current was not observed. Actin filaments disruption of VSM cells inhibits Ca_1 channel activity, whereas the microtubules disruption gave no effect on the activity. These results suggested that the maintenance of physiological function in VSM cells involve the functional-structural relationship between the Ltype Ca^ channel and cytoskeleton. The development of restenosis is dependent on the pathophysiological proliferation and then migration of the VSM cells could be prevented by control of L-type Ca^ channel activity and stabilization of the cytoskeleton in VSM cells., 論文

Changes in the cytoskeletal network alter the mechanical properties of cells that are essential for functions such as locomotion and cytokinesis. The transmembrane receptors and intracellular signals have been extensively studied on cytoskeletal changes in response to extracellular stimuli. After the procedure of percutaneous transluminal coronary angioplasty (PTCA) and highfrequency rotational atherectomy (PTCR) for stenosis of blood vessels, restenosis may be observed. The restenosis were generated under the proliferation and migration of vascular smooth muscle (VSM) cells. In addition, until recently it has been expected that the physiological regulation of VSM cells was dependent on L-type Ca^ (Ca_1) channel activity and structure of the cytoskeleton. To clarify the interactions between the structure of the cytoskeleton and activity of Ca_1 channel in VSM cell, we used electrophysiological and morphological experiments concomitantly. We investigated the effect of disruption of the actin filaments and the microtubules on Ca_1 channel activity (Ca^ current) of cultured VSM cells (A7r5 cell line) using whole cell voltage clamp. The results of immunostaining using anti-$ \alpha $-actin and anti-$ \alpha $-tubulin antibodies showed that colchicines disrupted both actin filaments and microtubules, while cytochalasin D and nocodazole disrupted only actin filaments, and microtubules respectively. Ca^ current was greatly reduced in cells that were exposed to colchicines or cytochalasin D but not to nocodazole. When the actin filaments were stabilized by phalloidine, the inhibition of Ca^ current was not observed. Actin filaments disruption of VSM cells inhibits Ca_1 channel activity, whereas the microtubules disruption gave no effect on the activity. These results suggested that the maintenance of physiological function in VSM cells involve the functional-structural relationship between the Ltype Ca^ channel and cytoskeleton. The development of restenosis is dependent on the pathophysiological proliferation and then migration of the VSM cells could be prevented by control of L-type Ca^ channel activity and stabilization of the cytoskeleton in VSM cells.

The type-I allergy is mediated by the interaction between IgE and its high affinity receptor (Fc$ \varepsilon $RI) followed by cross-linking with specific allergen. The binding site for the IgE was identified on the Fc$ \varepsilon $RI $ \alpha $ subunit, and the soluble $ \alpha $ subunit of Fc$ \varepsilon $RI was effective in controlling the lgE-mediated inflammation. Recent studies using IgE knock out mice revealed that IgG could also induce anaphylaxis. However, in the Fc$ \varepsilon $RI $ \alpha $ subunit knock out mice, IgE and IgG could not induce anaphylaxis. These results demonstrated that Fc$ \varepsilon $RI expression is essential for the development of the IgE-mediated inflammation. Further investigation on the interaction between IgE and Fc$ \varepsilon $Rl $ \alpha $ subunit on the membrane of target cells and on signal transduction in the cytoplasma and nucleus should offer novel concepts in developing strategies for the treatment of IgE-mediated inflammation.

A hybridoma producing monoclonal antibody against monoclonal DNP-specific rat IgE (IgE FE-3) was generated by fusion of P3-X63-Ag8-Ul (P3U1) mouse myeloma and spleen cells from BALB/c mice immunized with IgE FE-3. Five hybridoma clones were obtained. Three clones of them had a high reactivity against IgE FE-3 and a low reactivity against the L-chain of IgE FE-3 on ELISA analysis for the first screening of the hybridoma. The binding of monoclonal antibodies produced from clone BB-9 to IgE FE-3 revealed the strongest inhibition among three monoclonal antibodies on pretreatment with polyclonal rabbit anti-IgE FE-3 antibodies. It was clarified that monoclonal antibody BB-9 reacted specifically with the heavy chain of IgE FE-3 by Western blotting analysis. Furthermore, the reactivity of the monoclonal antibody BB-9 to IgE FE-3 was higher than that of MARE-1, which is a monoclonal antibody against rat myeloma IgE. The monoclonal antibody BB-9 was therefore employed for IgE-capture ELISA to determine the concentration of IgE FE-3. No significant alterations of color development occurred when igE FE-3 was concomitantly incubated with large excesses of IgG antibodies and rat myeloma IgE. Our data indicated that IgE-capture ELISA employing the monoclonal antibody BB-9 had a higher sensitivity for the determination of IgE FE-3 than did that using polyclonal rabbit anti-IgE FE-3 antibodies., 論文

To characterize the hemostatic disturbances after snakebite, we measured the antithrombin III (AT-III) activity, $ \alpha $_2-plasmin inhibitor ($ \alpha $_2-PI) activity, fibrinogen concentration (Fg) and fibrin degradation products (FDP) in 21 patients envenomed by several snakes in south China from August 1998 to October 1999. The hematological changes obtained were as follows: the mean activity of AT-III was decreased in patients bitten by Ophiophagus hannah (Oh.), Bungarus fasciatus (Bf.), Hydrophis cyanocinctus (Hc.), Rhabdophis subminiatus (Rs.), Trimeresurus stejnegeri (Ts.), the mean activity of $ \alpha $_2-PI was also decreased in all patients in the present study; Fg was not detectable in Rs. bite, while Fg concentration in Ts., Oh., He. and Bf. bites markedly decreased. The mean levels of FDP were therefore increased in all patients. Thus, it appears that DIC-like syndrome would be seen in patients envenomed by snakebites. In this study, we found that the patients who were bitten by Rs. which is still being classified as a non-venomous snake showed complete defibrinogenation syndrome with severe hemorrhage without any evidence of severe multiple organ damage. We also found that the patients of Ts. bite showed marked hemostatic disturbance without severe multiple organ damage as well as Rs. bite. It is thought that a discrepancy between hematological findings and clinical symptoms may be a characteristic phenomenon of DIC-like syndrome caused by snakebites, especially by Rs. and Ts. bite., 論文

The mechanism of invasion and metastasis of malignant tumors have been investigated in many reports especially on the blood coagulation-fibrinolysis system. Recently, on the other hand, it has been clarified that vascular endothelial growth factor (VEGF) plays important roles in tumor invasion and metastasis through the specific action on endothelial cells in tumor tissue. VEGF is known as an angiogenesis factor from tumor tissues, and it is thought to be an important endothelial mitogen for tumor growth; while, a vascular permeability factor (VPF) secreted from tumor tissues has been isolated, whose chemical and biological properties has been studied. Results show that VPF has been identified as VEGF; since VEGF increases vascular permeability and induces endothelial cell proliferation. VEGF also induces the expression of plasminogen activators, plasminogen activator inhibitor-1 or interstitial collagenase in endothelial cells. Platelet derived growth factor has proved to be an inducer of the transcription of VEGF gene by mediating protein kinase C. Hypoxia induces VEGF upregulation and protein expression, that could evoke the angiogenesis on tumor growth. Mutant ras oncogene upregulates VEGF expression. In addition, the expression of VEGF receptor on the endothelial cells in tumor tissues has increased. It has therefore been considered that the inhibitors of VEGF-induced angiogenesis should be useful materials for tumor therapy. Moreover, it has been found that antagonists against the VEGF-receptor, protein kinase C activity and blockers of VEGF-mediated signal transduction should be hopeful means for inhibition of tumor angiogenesis., 論文

We produced a monoclonal antibody against rabbit fibrinopeptide B (FPB) and established an immunological assay system to estimate it. Rabbit FPB was purified from the supernatant of fibrin clot prepared with rabbit fibrinogen and bovine thrombin. The purified rabbit FPB was coupled to keyhole limpet hemocyanin (KLH), and BALB/c mice were immunized with the resultant FPB-KLH. The spleen cells of immunized mice were hybridized with P3-X63-Ag8-U1 myeloma cells. As a result, one hybridoma (IA12/4A2) was selected, which secreted an antibody against rabbit FPB. The monoclonal antibody obtained belonged to the IgM class and its light chain consisted of a $ \kappa $-chain. The monoclonal antibody reacted not only with rabbit FPB but also rabbit fibrinogen, however it did not react with rabbit fibnnopeptide A. Utilizing this monoclonal antibody, we attempted to develop a competitive enzyme-linked immunoassay for estimating purified FPB. The working range of FPB varied between 5 and 50 $ \mu $g/ml.

Extracellular matrix of the ovary during ovulation is degraded and remodelled. After ovulation, angiogenesis and repair occur in the ovary. The dynamic change of extracellular matrix during ovulation is based on the role of proteolytic enzymes. Besides the fibnnolytic enzymes, the enzymes of collagenolytic system play important role in ovulation. In this article, we report that interstitial collagenase, type IV collagenase, collagenase inhibitor, tissue inhibitor of metalloproteinase 1 (TIMP-1) which belong to collagenolytic system play important role in the rupture of follicle, the migration of oocyte and the expulsion of oocyte from the ovary. During gonadotropin-induced ovulation, at 3 hr to 6 hr after hCG administration the interstitial collagenase mRNA increased, but at 9 hr after hCG administration TIMP-I mRNA and collagenase type IV mRNA increased in the granulosa cell, thecal compartment and stroma compartment. Both the interstitial collagenase and type IV collagenase were secreted in the latent form (procollagenase). This latent form, procollagenase was activated by fibrinolytic enzyme, plasmin and converted to active collagenase. It was clarified that the cross talk of fibrinolytic enzymes and collagenolytic enzyme has been observed in the process of ovulation., 論文

To study the hemostatic disturbances after snakebite, we measured the maximum platelet aggregation rate (MAR), antithrombin III (AT-III) activity, $ \alpha $_2 -plasmin inhibitor ( $ \alpha $_2 -PI) activity, fibrinogen concentration (Fg) and fibrin degradation products (FDP) in 17 patients with snakebite in South China from June 1996 to May 1997. The results obtained were as follows: 1) the mean MAR values in 15 samples from 13 patients before and without application of antivenom were markedly decreased in the case of snakebites from Vipera russelli (Vr.), Trimeresurus mucrosquamatus (Tm.) and Agkistrodon halys (Ah.); 2) the mean activities of AT-III were decreased in patients bitten by Deinagkistrodon acutus (Da.), Ah., Vr., Tm. and Naja naja atra (Nn.) but were not decreased in patients bitten by Trimeresurus stejnegeri (Ts.); 3) the mean activity of $ \alpha $_2-PI was decreased in all patients in the present study; 4) the mean concentration of fibrinogen (Fg) was markedly decreased in patients bitten by Da., Ah. and Vr., but was not decreased in the case of Nn. bite; and 5) the mean levels of FDP were increased in cases of Da.. Vr. and Ts. bite but not in Ah. and Tm. bite. After treatment using antivenom, the MAR, AT-III activity, $ \alpha $_2-PI activity and concentration of Fg in the patients with Nn. and Ah. bite were increased, but the FDP was not increased. In the case of patients who had been bitten by Vr. without treatment using antivenom, the hematological changes except for the AT-III activity did not return to normal levels within 45 hrs. The results of the present study indicate that disorders of platelet aggregation and the coagulation-fibrinolysis system are liable to occur in patients with snakebite from Da., Ah., Vr., Ts., Tm. and Nn. Furthermore, it appeared that DIC was evoked in some patients with these snakebites. Antivenom was found to be useful for improving the hemostatic disturbances after snakebite from Ah. and Nn., 論文

The animal facility affiliated with the School of Medicine, University of the Ryukyus, located in a subtropical area in Japan, was previously equipped with the COSA/TRON^【○!R】air recovery system in order to sustain a well balanced laboratory environment. In 1988, we measured the concentrations of ammonia, hydrogen sulfide, methyl mercaptan and dimethyl sulfide in the air of animal rooms at this animal facility. In this previous study, all of these odors were maintained at very low levels, a situation to which the COSA/TRON system was considered to have contributed. However, we anticipated that the air conditioning system would by now have become decrepit and deteriorated in its function for the elimination of malodorous substances and microorganisms. In the present study, we measured the same malodorous substances as those examined previously. The concentration of ammonia was determined by the indophenol method, and those of sulfur compounds by gas chromatography. The concentrations of ammonia, methyl mercaptan and dimethyl sulfide were increased as compared with those measured in 1988. The results obtained indicated that an increase in these malodorous substances (which could not be effectively eliminated by cleaning) had occurred, and that the COSA/TRON system continued to be useful for the elimination of sulfur compounds. However, efficiency of the system for eliminating ammonia at low concentrations may have gone down.

The role played by the fibrinolytic enzymes in blood circulation is well established. However, extravascular fibrinolysis has been observed in physiological and pathophysiological condition in the various organs. In this article the role of fibrinolytic enzymes in ovulation with regard to the rupture of the follicle and the significance of tissue type plasminogen activator (t-PA) derived from the granulosa cell is reviewed. It was clarified that two distinct PAs (t-PA, u-PA) and plasminogen activator inhibitor (PAI) were related to oyulation and the relationship between PAs and PAI was an important factor in ovulation. In addition, it was shown that gonadotropin and gonadotropin releasing hormone were closely related to the appearance of t-PA and PAI. It was emphasized that the fibrinolytic enzyme in the extravascular tissue played an important role in tissue remodelling, angiogenesis and repair besides ovulation., 論文

Rat immunoglobulin E (IgE) was purified from the serum of Wistar rats sensitized with DNP-Ascans. The IgE purification was carried out by precipitation with 50% saturated ammonium sulfate, ion-exchange chromatography on DEAE - Sephacel, affinity chromatography on goat anti-rat IgG (Fc) - Sepharose 4B and Protein A-Sepharose CL-4B, and gel filtration on Sephadex G-150. The results of the passive cutaneous anaphylaxis (PCA) reaction indicated that the protein fractionated by gel filtration on Sephadex G-150 was a rat IgE, and its molecular weight was approximately 130, 000 on SDS-PAGE analysis. The rat IgE obtained in the present study was designated as low molecular weight (LMW) rat IgE. The molecular weight of the LMW rat IgE corresponded to that of the myeloma IgE fragment (LMW rat myeloma IgE) produced by digestion of rat myeloma IgE (IR 162) with papain. The LMW rat IgE did not react with peroxidase (POD) - goat anti-rat myeloma IgE (Fc) on Western blotting. Conversely, LMW rat myeloma IgE reacted with POD-goat anti-rat myeloma IgE (Fc) on Western blotting. These findings indicate that structural differences between LMW rat IgE and LMW rat myeloma IgE may exist in the Fc region., 論文

We reviewed the characteristics and level of fibrinolytic enzymes in extracts of tissue with inflammatory disease and experimental inflammation in upper airway. In addition, we reviewed the changes of fibrinolytic activity in the circulating blood in both humans and experimental animals with acute inflammation. Tissue-type plasminogen activator (t-PA) existed in tissue extract of paranasal mucous membrane (PMM) with chronic sinusitis but urokinase-type plasminogen activator (u-PA) did not. Both t-PA and u-PA, however, existed in tissue extract of antrochoanal polyp (AP). The fibrinolytic activity in the circulating blood increased at the initial stage of human acute tonsillitis and Arthus tonsillitis of rabbits. It was presented that fibrinolytic enzymes in inflammation have an important role on extension, ending and repair at the local inflammatory sites. Furthermore, it was suggested that the suppression of the elevated activity of fibrinolytic enzyme might control the process of inflammation at the initial stage of acute inflammation., 論文

We evaluated the effect of administration of histamine-rat gammaglobulin conjugate (HRG) on the production of IgE in rats. The antigen-specific IgE level in the serum measured by PCA reaction decreased in the HRG-administered group, compared to that in the physiological saline-administered control group on day 15 after sensitization with DNP-Ascaris antigen. However, on day 10, HRG significantly increased the concentration of histamine compared to that in the control group. The IgG_1 level was higher in the HRG-administered group on day 15 and day 20, compared to that in the control group. From these results and the observation that the IgE production in rats increased to the highest level on day 15, we concluded that the decrease in PCA titer induced by the administration of HRG in rats was not dependent on an increase in IgE consumption, but on a decrease in IgE production. The increased histamine and IgG_1 level by the administration of HRG might have directly or indirectly suppressed the IgE production in rats.

We review the action of protease in airway inflammation from the standpoint of the protease-antiprotease system. The tracheobronchial secretion without inflammation has an abundant protease, plasminogen activator (PA) , which plays an important role in the continuous opening of the tracheobronchial lumina. Immediately after inflammation occurs in the airway tract, the protease activity is increased in the airway secretion, as compared to the protease activity in other organs. On the other hand, increased protease secretion, plays a strong role in the enlargement and exacerbation of the inflammation. In physiological inflammation, the increased protease activity is mediated and inhibited by protease inhibitor which is concomitantly elevated at the inflammatory locus. In inflammation of the airway tract, therefore, the application and supply of protease inhibitor to the airway tract, should cause the inflammation to subside. We consider that the role of protease inhibitor in limiting the inflammation of the airway tract is of clinical importance.

We developed a method for estimating tissue-type plasminogen activator (t-PA) antigen in tissue extract (TE) derived from nasopharyngeal carcinoma (NPC) by means of rocket immunoelectrophoresis (RIE). Initially, the TE sample without treatment failed to shape the immunoprecipitate line by RIE. However, when the TE sample was treated with final concentration of 0.2% SDS, we observed that the precipitate line was so clear that we could measure the height of the rocket., 論文