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本論文は，ダム貯水池の洪水調節機能向上を目的としダム放流量の新しい決定方法を提案するものである．著者らは従来から，現在まで降った降雨のうち確実にダム貯水池へ流入する量を，実際に流入する以前に放流する事前放流方法を提案している．本論文では，放流開始までの準備時間及び下流懸案地点における水位上昇速度を考慮してダムからの放流シミュレーションを行う．これにより各種法律，施設の放流能力，放流開始までの準備時間を考慮した実務に即した放流が可能な事を示すとともに，洪水低減効果向上のためには放流開始に要する準備時間を短くする必要がある事を示した．また，出水直前のダム貯水位が夏期制限水位以下の場合でも，初期水位低下量を考慮した事前放流を行うことで出水以降には夏期制限水位へと貯水位が回復することを明らかにした．

本論文は，物理的観点に立脚した降雨流出機構の解明および様々な流域に普遍的に適用可能な降雨流出計算手法の構築を目的とし，有効降雨に対して実測流量データから逆推定を行ったものである．実測流量データから求めた有効降雨と流域の保水能分布から求めた有効降雨の比較を行うことにより，有効降雨の時間変動特性および物理的解釈に対して考察を行った．これにより，有効降雨の物理的解釈は降雨の形で有効降雨をとる場合と流出寄与域の形で有効降雨をとるという二つのタイプを考える必要があることを示した．また，累積降雨量に応じた流出寄与面積，流出寄与斜面長の変動を考慮した流出計算手法を新たに提案し，草木ダム流域へ実際に適用することでその合理性・実用性を示した．

A new method of flood control by gate operation based on runoff characteristics of a basin is proposed in this paper. This method is based on an anticipatory release approach and the idea that there is no risk in reducing reservoir levels if the amount of anticipatory release is equal to the amount of inflow which will flow into the dam for certain from the rainfall that has already fallen. In this method, rainfall prediction or weather forecasting will not be necessary for flood control. The amount of anticipatory release will be calculated by the real time inflow into the reservoir or accumulated rainfall. And reservoir levels will certainly be restored to there former condition, because the amount of discharge is based on the area under the recession part of the hydrograph.

T cells play an important role in initiating autoimmune responses and maintaining synovial inflammation in rheumatoid arthritis. Although, anti-type II collagen antibody-induced arthritis (CAIA) is generally believed to be a T cell- and B cell-independent model, the detailed pathogenesis of CAIA remains unclear. In the present study, to elucidate the contribution of T cells to the pathogenesis of CAIA, we evaluated the effects of CTLA4 Ig and cyclosporin (CsA). Arthritis was induced in mice by intravenous injection of anti-type II collagen antibody followed by intraperitoneal injection of lipopolysaccharide. CTLA4 Ig was intraperitoneally administered and CsA was subcutaneously administered; then the severity of arthritis was evaluated by scoring the edema and erythema of paws and by measuring hind paw thickness. Paw samples were collected 12 days after the antibody injection, and the mRNA expression levels were analyzed by real-time quantitative polymerase chain reaction. Administration of CTLA4 Ig ameliorated the increases in arthritic score and paw thickness in the later phase, but not in the early phase of arthritis. CsA suppressed the increases in arthritic score and paw thickness in both the early and later phases of arthritis. CTLA4 Ig and CsA suppressed mRNA up-regulation of T-cell markers, CD3 and CD25, and immune response-related mediators, IFN-gamma and IL-12. They also suppressed the up-regulation of macrophage marker, F4/80, and proinflammatory cytokines, TNF-alpha, IL-1beta and IL-6. The results provide direct evidence that arthritis in this model is T-cell activation dependent.

Background/Aims: Phosphodiesterase type 4 (PDE4) has been previously shown to regulate colonic contractile activity in vitro. In this study, the effects of PDE4 inhibition were assessed in a model of stress-induced defecation previously demonstrated to be due to increased colonic transit/evacuation. Methods: Rats were individually placed in a mild restraint cage and placed into a 12°C environment (cold-restraint stress) for 60 min. Mice received restraint (only) stress at room temperature for 30 min. Loperamide (positive control compound) or two different PDE4 inhibitors (rolipram and roflumilast) were administered orally at several doses to the rodents 1 h before stress began. Vehicle alone was administered for comparison. The number of fecal pellets expelled during stress (fecal pellet output), total fecal pellet wet weight and total fecal water content were measured. Results: Loperamide produced a dose-related decrease (ID50s in mg/kg) in fecal pellet output (rat = 7.4, mouse = 0.7) and significantly decreased fecal wet weight (72.9%) and decreased fecal percent water content (9.4%). The two PDE4 inhibitors produced a similar dose-related inhibition of fecal pellet output. Rolipram exhibited ID50s in rat and mouse of 14.1 and 27.1, respectively. Rolipram significantly decreased fecal wet weight (58.8%) but increased fecal percent water content (15.0%). For roflumilast, ID50s were 24.2 mg/kg and 12.4 in the rat and mouse, respectively. Although roflumilast also significantly (p < 0.05) decreased fecal wet weight (47.2%), it did not significantly increase fecal percent water content. Conclusions: These data indicate that PDE4 inhibition is effective in reducing rodent stress-induced defecation, provides the first functional data on a potential role for PDE4 activity in the colonic evacuation response to stress, and indicates the potential utility of PDE4 inhibitors in functional bowel disease such as irritable bowel syndrome requires further evaluation.

Background: Recent advances in fibrosis biology have identified transforming growth factor (TGF)-β type I receptor-mediated activation of Smads as playing a central part in the development of fibrosis. However, to date, there have been few studies that examined the localisation and distribution of receptor-activated Smads protein (R-Smads: Smad2 and 3) during the fibrosis progression. Aims: To histopathologically assess the time-course change of the localisation and distribution of the Smads protein in pulmonary fibrosis. Methods: Pulmonary fibrosis was induced by intranasal injection of bleomycin (0.3 U/mouse). Lungs were isolated 2, 5, 7, 9 and 14 days after bleomycin treatment. Histological changes in the lungs were evaluated by haematoxylin-eosin stain or Masson’s trichrome stain, and scored. TGF-β1, Smad3 and phosphorylated Smad2 localisations in lung tissues were determined by immunohistochemistry. Results: The bleomycin treatment led to considerable pulmonary fibrotic changes accompanied by marked increase in TGF-β1 expression in infiltrating macrophages. With the progression in fibrosis (day 7–14), marked increases in Smad3-positive and pSmad2-positive cells were observed. There were intense Smad3-positive and pSmad2-positive signals localised to the nuclei of the infiltrating macrophages and to type II epithelial cells, and less intense signals in fibroblasts and hyperplastic alveolar/bronchiolar epithelial cells. Conclusions: The time-course data of TGF-β1 and R-Smads indicate that progressive enhancement of TGF-β1 signalling via R-Smad is activated in the process of fibrosis progression.

The purpose of present study is to clarify runoff mechanism and to understand hydraulic processes in mountainous basins. Considering multilayer flow in a slope, generation of overland flow for large scale flood is important. The method of multilayer runoff analysis which can calculate overland flow, subsurface flow and vertical infiltration flow based on soil property and rainfall form is newly proposed in this paper. Characteristics of overland flow in mountainous basins can be expressed rationally using our method. A method is applied to the Kusaki dam basin (254km2). The results of runoff analysis match well with the observed data in Kusaki dam basin and overland flow is expressed for peak time in a large scale flood.

Background: The regulatory mechanisms of IL-5 gene transcription in human peripheral T cells are unclear because the transfection efficiency of plasmid constructs into nontransformed T cells is very low. Methods: Concanavalin A (ConA)-stimulated blastocytes derived from peripheral blood lymphocytes of asthmatic subjects were transiently transfected with the human IL-5 gene promoter/enhancer-luciferase gene construct, pIL-5 (–511)Luc, and cultured with THP-1 cells (human monocytoid cells) and anti-CD3 monoclonal antibody (mAb). IL-5 level in the culture medium was determined by an enzyme-linked immunosorbent assay. Transcriptional activity of the IL-5 gene was measured by luciferase reporter analysis. Results: ConA-blast lymphocytes of asthmatic patients produced a significant amount of IL-5 upon combined stimulation with anti-CD3 mAb and THP-1 cells, but not with anti-CD3 mAb alone. Costimulation with anti-CD28 mAb also enhanced the anti-CD3 mAb- induced IL-5 production. Accordingly, luciferase activity induced by anti-CD3 mAb stimulation in pIL-5(–511)Luc-transfected ConA-blast lymphocytes was increased 1.9- and 3.4-fold by the addition of anti-CD28 mAb and THP-1 cells, respectively. Serial 5′ deletion analysis of the reporter gene demonstrated that the cis-regulatory element located at –119 to –80 is critical for anti-CD3 mAb-induced IL-5 gene transcription. Conclusions: Our present findings provide a useful model reflecting IL-5 gene transcription in human peripheral T cells in vivo, and clearly demonstrate that an interaction with monocytes enhances IL-5 gene transcription.

SummaryMechanisms that underlie the regulation of IL-5 gene expression in human peripheral T cells remain incompletely defined because of the low efficiency of transfection of plasmid constructs into non-transformed T cells. To elucidate the cellular and molecular mechanisms of IL-5 production, concanavalin A (ConA)-stimulated blastocytes derived from peripheral blood lymphocytes of asthmatic patients were employed in this study. Transcriptional activity of the synthetic human IL-5 promoter in ConA-stimulated blastocytes correlated with the production of IL-5. Deletion analysis of the reporter gene showed that the cis-regulatory element located at − 119 to − 80 is critical for inducible IL-5 promoter activity. Electrophoretic mobility shift assay revealed that an oligonucleotide probe corresponding to the element (− 119 to − 90) gave two specific bands. The slower migrating band was absolutely dependent on stimulation and was composed of a co-operative complex of the transcription factors, nuclear factor of activated T cells (NFAT) and activating protein-1 (AP-1). The faster migrating band was also inducible and was identified as AP-1-less NFAT. Mutation of either the NFAT or AP-1 element abrogated the slower migrating band and at the same time abolished transcriptional activity of the human IL-5 promoter/enhancer gene. Cyclosporin A equivalently suppressed DNA-binding activity of the composite NFAT/AP-1 site, promoter activity and protein production of IL-5. In conclusion, these data suggests that the composite NFAT/AP-1 binding element (− 115 to − 100) plays a crucial role in IL-5 synthesis by peripheral T cells of asthmatic patients.

The relationship between CD4(+) T cell-mediated airway eosinophilic inflammation and bronchial hyper-responsiveness (BHR) was investigated. Ovalbumin-reactive T(h)0 clones were adoptively transferred to unprimed BALB/c mice and then the mice were challenged by inhalation of the relevant antigen. Upon antigen provocation, infused T(h) clones infiltrated into the airways, followed by the accumulation and degranulation of eosinophils, goblet cell hyperplasia, edema and increase in bronchial responsiveness to acetylcholine. Transfer of several clones that differed in the levels of IL-5 production revealed that the magnitude of in vivo eosinophilia strongly correlated with the IL-5-producing capacity of the infused T(h) clones. Administration of anti-IL-5 mAb almost completely suppressed antigen-induced eosinophilic inflammation and BHR. Administration of anti-IL-4 mAb or anti-IFN-gamma mAb enhanced the eosinophilia and BHR, whereas anti-IL-2 mAb did not affect them. The number of accumulated eosinophils significantly correlated with the intensity of BHR. Our present results clearly demonstrated that CD4(+) T cells induced BHR as a result of eosinophilic inflammation. IL-5 totally regulated both responses.

Background: IL-5 plays a central role in allergic diseases associated with eosinophilic inflammation. We have previously reported that IL-5 production by peripheral blood mononuclear cells (PBMC) is greatly enhanced in both atopic and nonatopic asthmatics compared to control subjects. Method: Concanavalin A (Con A) blast lymphocytes were derived from PBMC of allergic and control subjects. Transcriptional regulation of the IL-5 gene was investigated by transient transfection assay. Results: A significant amount of IL-5 was produced by Con A blast lymphocytes derived from allergic subjects upon stimulation with phorbol ester and Ca2+ ionophore, whereas the cells derived from control subjects did not produce a detectable amount of IL-5. Production of IL-2 and IL-4 was not significantly different between the two groups. A luciferase reporter plasmid containing the human IL-5 promoter/enhancer region was transcribed by Con A blast lymphocytes derived from allergic subjects, but not by the cells from control subjects, upon activation. Conclusion: IL-5 synthesis by nontransformed T cells of allergic subjects is enhanced at the level of gene transcription. Elucidation of the molecular mechanism of IL-5 gene transcription by allergic T cells may delineate the pathogenesis of allergic disease for the future therapeutic intervention.

We examined the subtype-selective binding site of the beta-adrenergic receptors (betaARs). The beta(1)/beta(2)-chimeric receptors showed the importance of the second and seventh transmembrane domains (TM2 and TM7) of the beta(2)AR for the binding of the beta(2)-selective agonists such as formoterol and procaterol. Alanine-substituted mutants of TM7 of the beta(2)AR showed that Tyr(308,) located at the top of TM7, mainly contributed to beta(2) selectivity. However, Tyr(308) interacted with formoterol and procaterol in two different ways. The results of Ala- and Phe-substituted mutants indicated that the phenyl group of Tyr(308) interacted with the phenyl group in the N-substituent of formoterol (hydrophobic interaction), and the hydroxyl group of Tyr(308) interacted with the protonated amine of procaterol (hydrophilic interaction). In contrast to beta(2)AR, TM2 is a major determinant that beta(1)-selective agonists such as denopamine and T-0509 bound the beta(1)AR with high affinity. Three amino acids (Leu(110), Thr(117), and Val(120)) in TM2 of the beta(1)AR were identified as major determinants for beta(1)-selective binding of these agonists. Three-dimensional models built on the basis of the predicted structure of rhodopsin showed that Tyr(308) of the beta(2)AR covered the binding pocket formed by TM2 and TM7 from the upper side, and Thr(117) of the beta(1)AR located in the middle of the binding pocket to provide a hydrogen bonding for the beta(1)-selective agonists. These data indicate that TM2 and TM7 of the betaAR formed the binding pocket that binds the betaAR subtype-selective agonists with high affinity.

1. To delineate the mechanism by which cyclic AMP (cAMP) suppresses interleukin (IL)-5 synthesis, the effects of prostaglandin (PG) E2, forskolin, dibutyryl (db)-cAMP and the Ca2+ ionophore, ionomycin on cytokine synthesis, proliferation and CD25 expression of human T cells were investigated. Further studies were performed by measurement of the intracellular concentrations of cyclic AMP ([cAMP]i) and Ca2+ ([Ca2+]i) and by electrophoretic mobility shift analysis (EMSA). 2. PGE2, forskolin and db-cAMP suppressed IL-5 production by human T cell line following T cell receptor (TCR)-stimulation. PGE2 suppressed TCR-induced messenger RNA (mRNA) expression of IL-2, IL-4 and IL-5, as well as proliferation and CD25 expression. 3. Cyclic AMP-mediated suppression of cytokine synthesis, proliferation and CD25 expression in human T cells were attenuated by ionomycin. 4. [cAMP]i was increased by PGE2 and forskolin. PGE2 suppressed the TCR-induced biphasic increase in [Ca2+]i. EMSA revealed that four specific protein-DNA binding complexes related to NF-AT were detected at the IL-5 promoter sequence located from -119 to -90 relative to the transcription initiation site. The slowest migrating complex induced by TCR stimulation was enhanced by PGE2 and further upregulated by ionomycin. Another binding which did not compete with cold AP-1 oligonucleotides, was constitutively present and was unaffected by PGE2 but enhanced by ionomycin. 5. The suppressive effect of cyclic AMP on human IL-5 synthesis is mediated by interference with intracellular Ca2+ mobilization but distinct from the NF-AT-related pathway.

Background: We have recently demonstrated that allergic eosinophilic inflammation is transferred to unprimed mice by infusing IL-5-producing CD4+ T cells. The contribution of mast cells to the development of eosinophilic inflammation is controversial. Methods: To clarify the possible different roles of CD4+ T cells and mast cells in eosinophilic inflammation, we compared antigen-induced airway eosinophilia between mast-cell-deficient mice (WBB6F1-W/Wv) and their congenic normal littermates (WBB6F1-+/+). Results: The time course study indicated that equivalent numbers of eosinophils were recruited into the airway of both +/+ and W/Wv mice 6, 24, 96, and 216 h after antigen challenge, whereas the number of eosinophils 48 h after antigen challenge was significantly lower in W/Wv compared to +/+ mice. Administration of either anti-CD4 or anti-IL-5 monoclonal antibody almost completely inhibited antigen-induced eosinophil recruitment in W/Wv mice 48 h after antigen challenge. In contrast, the inhibitory effect of these antibodies in +/+ mice were partial (∼50% inhibition). Anti-CD4 and anti-IL-5 antibodies equally suppressed airway eosinophilia in both +/+ and W/Wv mice 96 h after antigen challenge. Conclusion: Our study indicates that CD4+ T cells are crucially involved in the development of allergic eosinophilic inflammation, while mast cells may play a supplemental role depending on the kinetics of the response.

1 We have recently demonstrated that airway eosinophilic inflammation can be transferred to unprimed mice by infusing interleukin (IL)-5-producing T cell clones. Using that murine model, we performed this study to delineate the mechanism of cyclosporin A and dexamethasone to inhibit allergic airway eosinophilic inflammation. 2 The ovalbumin-reactive murine T cell clones, FJ17, produced IL-2, IL-4 and IL-5 upon stimulation with relevant antigen. In FJ17-transferred mice, messenger RNA (mRNA) of IL-2 and IL-5 expressed in the lungs, the number of eosinophils in bronchoalveolar lavage fluid (BALF) was increased and the bronchial responsiveness to acetylcholine was enhanced after antigen provocation. 3 Cyclosporin A (10, 100 ng ml−1) and dexamethasone (10, 100 ng ml−1 suppressed the production of IL-5 as well as IL-2 and IL-4 by FJ17 in vitro. 4 Subcutaneously administered cyclosporin A (30 mg kg−1) and dexamethasone (10 mg kg−1) inhibited antigen-induced mRNA expression of IL-2 and IL-5, increase of BALF eosinophils and bronchial hyperresponsiveness of FJ17-transferred mice in vivo. The number of BALF eosinophils was correlated with the bronchial responsiveness to acetylcholine (r = 0.672). 5 The results clearly indicated that the suppression of IL-5 synthesis by T cells is involved in the effects of cyclosporin A and dexamethasone to inhibit allergic airway eosinophilic inflammation. British Journal of Pharmacology (1998) 124, 1425–1432; doi:10.1038/sj.bjp.0701982

Effect of a selective type 4 phosphodiesterase (PDE4) inhibitor, T-440, on intracellular cyclic AMP (cAMP) level and interleukin (IL)-2 production of Jurkat cells was investigated. T-440 suppressed both cAMP-PDE activities in cytosolic and membrane fractions of Jurkat cells. Intracellular cAMP level in Jurkat cells was elevated by PGE2 and forskolin but not by T-440. T-440, however, significantly enhanced the increase of cAMP by PGE2. PGE2 and forskolin inhibited IL-2 production of Jurkat cells stimulated with concanavalin A. T-440 by itself did not affect IL-2 production, but significantly enhanced the effect of PGE2 on IL-2 production. The increase of intracellular cAMP by T-440, PGE2, forskolin and T-440 plus PGE2 was well correlated with the inhibition of IL-2 production. These results indicate that IL-2 production of T cells is regulated by cAMP-PDE activity. Immunomodulatory effects of PDE4 inhibitors like T-440 should further be explored in vivo.

Background: The Lewis rat among highly inbred strains exhibits significant airway hyperresponsiveness (AHR) following intravenous administration of Sephadex G-200 (Sephadex). The aim of this study was to investigate the association of Sephadex-induced AHR with changes in airway inflammation in Lewis rats. Methods: A suspension (0.5 mg/ml/rat) of Sephadex was intravenously administered to male Lewis rats on days 0, 2 and 5. Measurement of airway responsiveness to serotonin, bronchoalveolar lavage (BAL) and histological study were performed on day 2–11. Results: Significant AHR induced by Sephadex was recognized on day 2 (p < 0.05), and AHR reached a maximum on day 7 (p < 0.001). In the BAL study, eosinophils increased on day 2 (p < 0.01) with a peak on day 5 (p < 0.05). In the histological study, we found Sephadex beads trapped in small arteries of the lung and granulomatous arteritis on day 2 or later. Pulmonary granulomas, horseshoe-shaped multinuclear giant cells, eosinophils and goblet cell hyperplasia were observed on day 2, and the degree became intense on day 5–7. GCC-AP0341 (10 mg/kg, i.p. × 3) inhibited the recruitment of eosinophils in BAL fluid and in lung tissue, but it did not inhibit AHR. The compound also inhibited pulmonary granulomas and goblet cell hyperplasia. Conclusion: The mechanism of Sephadex-induced AHR may not be directly associated with inflammatory changes such as recruitment of eosinophils, pulmonary granulomas and hyperplasia of goblet cells in rats.

We investigated the effects of dexamethasone and cyclosporin A on Sephadex bead (Sephadex G-200, Sephadex)-induced airway hyperresponsiveness (AHR) and inflammation in rats. Sephadex (0.5 mg/animal) was intravenously administered on days 0, 2 and 5. Bronchoalveolar lavage, histological study and measurement of AHR were performed on day 7. Dexamethasone (0.1, 1 and 10 mg/kg, p.o. × 3) and cyclosporin A (0.1, 1 and 10 mg/kg, s.c. × 3) clearly inhibited the increase in eosinophils in bronchoalveolar lavage fluid after Sephadex injection. On histological study, pulmonary eosinophilia, granulomatous arteritis with horseshoe-shaped multinuclear giant cell formation and goblet hyperplasia were observed after Sephadex injection. Both dexamethasone (10 mg/kg x 3) and cyclosporin A (10 mg/kg x 3) inhibited these findings and an increase in eosinophil peroxidase in the lung. Dexamethasone dose-dependently inhibited AHR induced by Sephadex, and completely suppressed it at a dose of 1 mg/kg (× 3). Cyclosporin A, however, did not inhibit AHR even at a dose of 10 mg/kg (× 3). These results show that there is a difference between dexamethasone and cyclosporin A in the inhibitory effect on Sephadex-induced AHR, and they suggest that eosinophils are not directly associated with the development of AHR after Sephadex injection.

1. We have studied the difference in receptor binding activity between partial and full beta2-adrenoceptor agonists and the abilities of the agonists to interact with Ser204 and Ser207 in the fifth transmembrane region of the beta2-adrenoceptor, amino acid residues that are important for activation of the beta2-adrenoceptor. 2. In the binding study with [125I]-iodocyanopindolol, the Ki values of (+/-)-salbutamol, (+/-)-salmeterol, TA-2005 and (-)-isoprenaline for the beta2-adrenoceptor expressed in COS-7 cell membranes were 3340, 21.0, 12.0 and 904 nM, respectively. The beta1/beta2 selectivity of these agonists was in the order of (+/-)-salmeterol (332 fold) > TA-2005 (52.8) > (+/-)-salbutamol (6.8) > (-)-isoprenaline (1.1), and the beta3-/beta2-adrenoceptor selectivity of these agonists was in the order of TA-2005 (150 fold) > (+/-)-salmeterol (88.6) > (+/-)-salbutamol (10.4) > (-)-isoprenaline (3.2). 3. The maximal activation of adenylyl cyclase by stimulation of the beta1-, beta2- and beta3-adrenoceptors by TA-2005 was 32, 100 and 100% of that by (-)-isoprenaline, respectively, indicating that TA-2005 is a full agonist at the beta2- and beta3-adrenoceptors and a partial agonist at the beta1-adrenoceptor. (+/-)-Salbutamol and (+/-)-salmeterol were partial agonists at both beta1- (8% and 9% of (-)-isoprenaline) and beta2- (83% and 74% of (-)-isoprenaline) adrenoceptors. 4. The affinities of full agonists, TA-2005 and (-)-isoprenaline, were markedly decreased by substitution of Ala for Ser204 (S204A) of the beta2-adrenoceptor, whereas this substitution slightly reduced the affinities of partial agonists, (+/-)-salbutamol and (+/-)-salmeterol. Although the affinities of full agonists for the S207A-beta2-adrenoceptor were decreased, those of partial agonists for the S207A-beta2-adrenoceptor were essentially the same as for the wild type receptor. 5. The constitutively active mutant (L266S, L272A) of the beta2-adrenoceptor had an increased affinity for all four agonists. The affinities of full agonists were decreased by substitution of Ser204 of the constitutively active mutant, whereas the degree of decrease was smaller than that caused by the substitution of the wild type receptor. Although the affinities of (+/-)-salbutamol and (+/-)-salmeterol for the S207A-beta2-adrenoceptor were essentially the same as those for the wild type beta2-adrenoceptor, the affinities of (+/-)-salbutamol and (+/-)-salmeterol for the constitutively active beta2-adrenoceptor were decreased by substitution of Ser207. 6. These results suggest that Ser204 and Ser207 of the wild type and constitutively active beta2-adrenoceptors differentially interacted with beta2-selective agonists.

We investigated the effects of phosphodiesterase (PDE) inhibitors on ozone-induced airway hyperresponsiveness (AHR) in guinea pigs. Theophylline (10–100 mg/kg), rolipram (1–100 mg/kg) and T-440 (1–100 mg/kg) were orally administered 30 min before ozone exposure (3 ppm, for 30 min). Ozone exposure caused an increase in airway responsiveness to methacholine aerosol, and log PC 10 (log-transformed methacholine concentration causing a 10 cm H 2 O increase in pulmonary inflation pressure) in the control and ozone-exposed group was 4.43±0.05 ( n =6) and 3.26±0.15 ( n =12), respectively. Theophylline at 100 mg/kg, 10 mg/kg rolipram and 10 mg/kg T-440 significantly inhibited AHR with log PC 10 value of 4.73±0.16 ( n =7), 3.74±0.11 ( n =7), 3.82±0.15 ( n =6), respectively. On histological examination, epithelial damage in the trachea and peripheral airways was recognized after ozone exposure. At 100 mg/kg, rolipram, T-440 and theophylline caused complete inhibition of AHR, and prevented epithelial damage of the trachea and peripheral airways. These results indicate that PDE inhibitors prevent not only ozone-induced AHR but also airway epithelial injury by ozone.

Bronchial asthma is characterized by chronic eosinophilic inflammation of the bronchial mucosa. Accumulating evidences suggest that activated T cells and T cell cytokines play critical roles in the local accumulation and activation of eosinophils. To further delineate the critical role of T cells on asthma, we tested the possibility whether eosinophilic inflammation of the bronchial mucosa is induced by transferred T cell clones, in the absence of antigen-specific immunoglobulins (IgE, A, and G). Ovalbumin-specific Th2 clones were established and cytokine profiles were determined. Eosinophilic inflammation accompanied with airway hyperresponsiveness occurred only when unprimed mice were transferred with IL-5 producing Th2 clones and challenged by the inhalation of relevant antigen. Increase of IL-5 concentration in bronchoalveolar lavage fluid (BALF) was detected after the challenge, indicating the local production of cytokines by the transferred T cells, and preceded the appearance of the airway eosinophilia. Eosinophil infiltration was completely suppressed by the administration of anti-IL-5 neutralizing antibody, indicating the essential role of IL-5 in this model. The intensity of the eosinophil accumulation in vivo correlated well with the capacity of the T cell clones to produce IL-5 in vitro. We concluded that the existence of IL-5-producing helper T cells is sufficient for the development of the eosinophilic inflammation at the bronchial mucosa upon inhalation challenge of the relevant antigen.

蛇行河川の湾曲部においては, 遠心力に起因する二次流が生じ, 外岸付近の河床の洗掘や側岸の浸食が起き, これが河川管理上重要な問題となっている. この対策として、本研究では、桟型粗度を側壁上に, 上端を下流側に倒すような形で斜めに設置することを考えた. 本研究では, まず, 直線水路における広範囲にわたる実験的検討を通して, 斜め桟粗度により生成される流れ場の諸特性を明らかにしている. 次に, 斜め桟粗度を湾曲部外岸側壁に設置することによって, 遠心力による二次流を人工的に制御でき, 外岸付近の局所洗掘および側壁浸食を抑制できることを明らかにしている. さらに, 本方法を実河川に適用するに当たって必要となる桟粗度の最適配置法についての基礎的な検討を行った.

We investigated the selectivity of T-440 for the inhibition of phosphodiesterases (PDEs) in vitro and for bronchial anti-spasmogenic effects in vivo. Using a fast protein liquid chromatography system, five PDE isozymes, PDE 1, PDE 2, PDE 3, PDE 4 and PDE 5 were prepared from guinea pig and dog tissues. T-440 selectively inhibited PDE 4 with an IC50 of 0.071 microM and 0.13 microM for guinea pig lung and dog trachea, respectively. The IC50 values for all other PDE isozymes were over 20 microM. In contrast, theophylline nonselectively inhibited all the tested PDE isozymes, and the inhibition did not exceed 50%, even at 100 microM. T-440 inhibited the histamine-induced bronchoconstriction of anesthetized dogs in a dose-dependent manner with an ED50 of 0.029 mg/kg, indicating that T-440 is 600 times more potent than theophylline. Both T-440 and theophylline increased LV dp/dt/P (LVP; left ventricular pressure) in anesthetized dogs with ED50 values of 3.6 mg/kg and 4.4 mg/kg, respectively. This potency was 1/125 times the bronchial anti-spasmogenic effects for T-440 and 4.2 times that of theophylline. Rolipram, a PDE 4 inhibitor, also showed selective bronchial anti-spasmogenic effects in anesthetized dogs. These results suggest that T-440, which specifically inhibits PDE 4 activity, has potent and selective bronchial anti-spasmogenic effects.

Type IV phosphodiesterase (PDE IV) is expressed in many tissues including airway smooth muscle and inflammatory cells, suggesting that this isozyme has a regulatory role in the pathogenesis of bronchial asthma. We investigated the effect of a novel selective PDE IV inhibitor, T-440, on immediate asthmatic reaction (IAR), late reaction (LAR) and leukocyte infiltration into the airways after allergen challenge in guinea pigs. Inhalation of ovalbumin by sensitized guinea pigs induced an immediate increase in specific airway resistance that peaked at 15 min. LAR was only produced in combination with chronic treatment with metyrapone, an inhibitor of the biosynthesis of adrenocortical steroids, and was suppressed by administration of dexamethasone, suggesting that the expression of LAR is dependent on an intrinsic steroid hormone. These reactions were accompanied by increases in the number of eosinophils and neutrophils recovered in bronchoalveolar lavage fluid. Oral administration of T-440 significantly inhibited IAR, LAR and the infiltration of eosinophils, whereas it suppressed neutrophil accumulation with relative low potency. These findings imply that the inhibition of PDE IV activity is a reasonable target for the management of obstructive airway diseases associated with airway inflammation such as asthma.

Structual basis of the β-adrenergic receptor (βAR) to confer the β selectivity is largely unknown. We investigated the binding regions of βAR which is important for the subtype selective binding. TA-2005 is a highly selective β2 agonist with the carbostyril structure. Binding regions of the βAR for TA-2005 was investigated by use of chimeric β1 and β2 receptors as comapared with those of non-selective β agonist isoproterenol. Replacement of transmembrane region 2 (TM2) or TM7 of β2AR with those of β1AR decreased the affinity of TA-2005. Deletion of methoxyphenyl group from TA-2005 molecule lost β2 selectivity. Then it is reasonable to assume that methoxyphenyl group interacts with TM2 and TM7 of β2AR, and contributes to the β2 selectivity. Several β2 selective agonists including formoterol and salmeterol also required TM2 or TM7 for the β2 selective binding. T-0509 is a β1 selective full agonist. In contrast with the binding of β2 selective agonist, selective binding of T-0509 to β1AR required the only TM2 of β1AR. These data suggested that TM2 and TM7 of βAR is important for the subtype selective binding.

Revetments are constructed to reduce erosion of river banks, but it is very difficult to design hydraulically. In this paper, hydraulic properties are investigated experimentally. As the result, squared bars which are used for the roughness of river banks will prevent the erosion at the concave side and will cancell the circulation in sinuous rivers.

Background: The crucial role of T helper (Th) 2 cells in allergic eosinophilic inflammation has been established. Methods: To determine the dynamics of Th2 cells in airway eosinophilic inflammation, ovalbumin-reactive murine Th2 clones were infused into unprimed mice. Results: The infused T cells were activated and migrated into the bronchial submucosa upon provocation with the relevant inhaled antigen. The T cell migration was followed by massive eosinophilic infiltration, edema, swelling of epithelial cells in the lung and a significant increase in bronchial responsiveness to acetylcholine. Conclusion: Clonal Th2 cells have the potential to accumulate and induce eosinophilic inflammation in the lung upon antigenic stimulation.

A series of 1-aryl-2,3-bis(hydroxymethyl)naphthalene lignans have been synthesized and evaluated for their ability to selectively inhibit PDE IV isolated from guinea pig. Replacement of the 1-phenyl ring by a pyridone ring led to marked improvement of their selectivity for PDE IV over PDE III. The compounds that were most potent and selective involved those bearing an N-alkylpyridone ring at C-1. These compounds also showed potent antispasmogenic activity without causing significant changes in heart rate in the guinea pig. The most potent compound was 6,7-diethoxy-2, 3-bis(hydroxymethyl)-1-[1-(2-methoxyethyl)-2-oxo-pyrid-4-yl]nap hth alene (17f), ED50 values of histamine-induced and antigen-induced bronchoconstriction in the guinea pig being 0.08 and 2.3 mg/kg iv, respectively. This compound was chosen as a candidate for further pharmacological evaluation.

都市河川の河床上に堆積した有機物などがもたらす水質問題の対策に, エアリフトと呼ばれる散気装置を利用することを考えた. 本論文は, 水質改善のためにエアリフトの水理機能を検討したものであり, エアリフトによる揚水流量の計算法と循環流が効率よく生じるアスペクト比を明らかにした. また, 現地実験を行い, エアレ-ションによるDO濃度の増加と底泥浮上によるSS濃度の増加の測定結果から水質改善機能を確かめた.

A numerical model of saltation akin to those of previous researchers is presented to investigate the detailed nature of bed load motion in terms of the mechanics of saltation. The model is deterministic in the computation of particle trajectory, but probabilistic in terms of bed collision. This probabilistic element arises from the random structure of the bed. In the present analysis, a stochastic treatment of collision allows for a fairly complete model of saltation in water. Characteristic quantities, including the sediment transport rate itself, are derived and compared with experimental data. The model yields good agreement with data with a minimum of assumptions. The assumed three-dimensional random structure of the bed is a unique feature of the present model. It allows for the prediction of several phenomena that cannot be obtained via the assumption of a regular two-dimensional bed structure.

Bronchial asthma is an inflammatory disease of the airways. The Sec14l3 gene, encoding a 45-kDa secretory protein, is specifically expressed in airway epithelium. Here, we report on the kinetics of Sec14l3 expression following allergic inflammation of the lung. Brown Norway rats were sensitized by intraperitoneal injection of ovalbumin, followed by challenge with aerosolized ovalbumin after a 3-week interval. This animal model showed many features similar to human allergic asthma: an increase in inflammatory cells such as eosinophils, lymphocytes and neutrophils in bronchoalveolar lavage (BAL) fluid and histopathological alteration of lung tissue, exhibiting infiltration of these inflammatory cells and degeneration and necrosis of alveolar epithelium. These parameters reached their maximal level 24h after allergen challenge. In contrast, quantitative polymerase chain reaction analyses demonstrated a rapid and significant reduction of Sec14l3 mRNA in lung tissue and maximum reduction (to 1.4% of the control) was observed at 24h. Pretreatment with dexamethasone significantly suppressed both the Sec14I3 mRNA reduction and all of the inflammatory changes. The 45-kDa secretory protein was identified in the supernatant of BAL fluids. Two-dimensional gel images of the supernatant proteome also revealed down-regulation of the protein following inflammation (to approximately 30% of the control at 24h). Thus, Sec14l3 expression is highly and inversely associated with the progression of airway inflammation. Sec14l3 mRNA and protein may function in the homeostasis of airway epithelial cells under normal conditions.

Temporary cofferdam used in river improvement work causes the backwater. In flood time, this backwater increases the risk of overflow over the bank. To diminishthis backwater there are two method. One is kinematic method which is to diminish the flow resistance by using smooth shutterings in shape and rough-ness. The other is dynamic method which is to diminish this backwater by adding momentum ofaccelerator. We carried the mock-up test of dynamic method using the propellers of work-ship as accelerator, and as the result the water level fall down 10 cm. The result exhibited good agreement with the momen-tum principle. This paper presents the result of mockup test.

Tracheal relaxing effects and beta 2-selectivity of TA-2005 were investigated by functional experiments and radioligand binding assay in guinea pigs in comparison with those of other beta-agonists, isoproterenol, procaterol, formoterol and salbutamol. The relaxing activity of TA-2005 on histamine-induced contraction in the isolated trachea was most potent among the five agonists, and it was blocked by a beta 2-selective antagonist (ICI 118,551) but not by a beta 1-selective antagonist (bisoprolol). The potency of the relaxing effect was in the order of TA-2005 (pD2 = 9.79) greater than formoterol greater than procaterol greater than isoproterenol greater than or equal to salbutamol. The positive chronotropic effect of TA-2005 was similar to that of isoproterenol; and it was more potent than those of formoterol, procaterol and salbutamol in the isolated atria. The selectivity for tracheal muscle to atria of these agonists were in the order of procaterol greater than greater than or equal to formoterol greater than TA-2005 greater than salbutamol much greater than isoproterenol. A radioligand binding experiment using guinea pig lung and cardiac ventricle as beta 2- and beta 1-adrenoceptor sources, respectively, has also demonstrated that TA-2005 possesses extremely high affinity (IC50 = 1.04 nM) and selectivity (38-fold) to beta 2-adrenoceptors. By addition of GTP, the competition curve of [125I]iodocyanopindolol shifted rightward, indicating the agonist property. These results confirmed that TA-2005 is a highly beta 2-selective agonist that exerts a potent tracheal relaxing effect.

The flow in an open channel with the non-uniformly sloping bed represents the complicated flow pattern, and the feature of the flows are made clear by the experiments. The following 3 types of flows can be observed: I) the super-critical flow, II) the subcritical flow, and III) the transitional flow with the undular jump. A distinct inner turbulent boundary layer can be observed for each type of flow.It is important to analyze the flow pattern in the case II) practically. The velocity distribution near the bed follows the 1/n-th power law even in such cases. The bed shear stress can be estimated by turbulent boundary layer equation, and the distribution of vorticity along the stream bed can be estimated by using the power law of velocity distribution.Consequently, the numerical simulation model based on the above facts is proposed by using the rotational form of Navier-Stokes equation.

The purpose of this paper is to study the charactristics of fluid forces acting on spherical roughness elements which placed densely and sparsely. By measuring the velocity distribution above the roughness elements, it is possible to determine the shearing stress on the rough wall with the aid of the logarithmic formula and hence the equivalent roughness length. On the other hand, Drag Force and Lift Force were calcurated from the surface pressure of the roughness elements which were measured directly. The integrated mean shear values have been compared with those estimated from directry measured Drag Force and the energy gradient.

The purpose of this experimental study is to measure the fluid force acting on the square timber laid on a revetment. By the orifice installed through the square timber, the heads of fluid, surface pressure, are measured to calculate the drag force and shearing stress. It's ascertained the drag coefficient CD is almost constant on it throughout entire span. So, it's possible to estimate the drag force and flow resistance by determining the drag coefficient.

A convex or concave boundary of the flow cross section causes a three-dimensional flow.Such a three-dimensional flow has an effect on flow structure which is very difficult to deal with. As the first step to study, a flow charastatic of variation due to the shape of cross section and Reynolds number is clarified experimentally.Furthermore, in consideration of its practical application to rivers, the hydraulic characteristic of compound channel with floodplane or berm of levee is studied.

The availavility of the ω-ψ method to simulate the flow field in an open channel with gradually varied bed slope was studied. But the eddy viscosity, included in the simulation and very important factor in hydraulics, could not be well-considered at that stage, since, for such flow, the boundary condition was dominant rather than the eddy viscosity. In this paper, the fundamental feature of that factor is studied through the simulation of the two-dimensional boundary layer flow on the flat plate in an open channel. For the turbulent boundary layer flow, the relation between eddy viscosity as a mean value of the flow field and mesh size or Reynolds number is made clear. On the other hand, for the laminar flow, the apparent relation among such factors is not found.

The effect of a novel anti-allergic drug, betotastine besilate (betotastine) on interleukin (IL)-5 production by human peripheral blood mononuclear cells (PBMC) was investigated. PBMC of Dermatophagoides farinae extract (Df)-sensitive donors produced IL-5 and showed a proliferative response upon stimulation with relevant antigen (10 microg/ml). Df-induced IL-5 production by PBMC was significantly inhibited by betotastine at 10 and 100 microM. Betotastine also suppressed proliferation of PBMC with less potency. The effect of betotastine on IL-5 production was enhanced and significant even at 0.1 microM when the drug was added 120 min before antigen stimulation. Ketotifen and cetirizine also inhibited IL-5 production, but the effects of these drugs were significant only at 100 microM. These findings indicate that the suppression of IL-5 production may be involved in the anti-allergic effect of betotastine.

Cathepsin K is a cysteine proteinase that is highly expressed by osteoclasts and is being pursued as a potential drug target for the treatment of osteoporosis. We have reported that microcomputed tomography (micro-CT) analysis of bone microarchitecture may serve as a valuable tool for evaluating both antiresorptive and anabolic agents in ovariectomized (OVX) mice. The purpose of this study was to evaluate the effect of SB-553484, a novel cathepsin K inhibitor (human Ki,app=0.14 nM, mouse Ki,app=26 nM), on the OVX mice by micro-CT bone morphometric analysis. Seven weeks female BALB/c mice were OVX or sham-operated. OVX animals were treated with SB-553484 (30 mg/kg, sc) or Rolipram (10 mg/kg, po), a phosphodiesterase 4 inhibitor used as a positive bone anabolic agent, twice a day for 2 weeks. Both SB-553484 and Rolipram significantly prevented the decrease of trabecular bone volume as well as the deterioration of trabecular architecture in OVX mice. Interestingly, SB-553484 demonstrated a more pronounced effect in improvement of trabecular separation, number and connectivity, and a weaker effect in improvement of trabecular thickness compared to that of Rolipram. These differences indicate that SB-553484 mainly acted as an antiresorptive agent in OVX-induced loss of trabecular bone. On the other hand, SB-553484 significantly increased cortical bone volume and cortical thickness as well as Rolipram in OVX mice indicating an unexpected stimulatory effect of SB-553484 on cortical bone formation. These data suggest that targeting cathepsin K may prove therapeutically beneficial in the treatment of diseases with accelerated bone loss such as postmenopausal osteoporosis not only by inhibiting bone resorption but also by potentially stimulating cortical bone formation.

OBJECTIVES The purposes of this study were to develop an osteoporosis model in a short period of 2 weeks after ovariectomy in mice and to investigate whether analysis of microcomputed tomography (muCT) 3-dimensional bone parameters could provide useful information on the mechanism of action of antiosteoporotic agents. MATERIALS AND METHODS Mice were ovariectomized (OVX) or sham-operated, and the OVX mice were treated daily with 17beta-estradiol (E2), parathyroid hormone (PTH[1-34]), raloxifene, rolipram, or vehicle for 2 weeks. On day 14 post-OVX, the left femur bones were removed and then the distal metaphyseal bone was analyzed by both muCT and histomorphometry. RESULTS The trabecular bone volume, thickness, number, and connectivity significantly decreased and the number of osteoclasts increased in OVX mice. Treatment of OVX animals with each of the 4 antiosteoporotic agents significantly increased the bone volume and improved the bone architecture. However, the improvement of trabecular thickness in the rolipram-treated group and that of cortical thickness in the PTH(1-34)-treated group were the most marked, whereas the improvement of connectivity in the rolipram-treated group was the least among the drug-treated groups. These different improving effects of agents on the bone parameters reflect the differential effects of these agents on bone formation and bone resorption. CONCLUSIONS This study demonstrated the feasibility of evaluating the effect of the antiosteoporotic agents within 2 weeks after ovariectomy in mice. The muCT analysis may serve as a valuable tool, specifically in a high-throughput pharmacological screening test, offering useful information regarding the effects of test compounds on both bone resorption and formation.