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The cell-volume regulation by swelling-activated Cl- current (I(Cl,swell)) was studied in guinea pig ventricular myocytes, using a microscopic video-image analysis. We have previously shown that in ventricular cells depolarized in high-K+ ([K+]o>45 mM) solution, an activation of the cyclic AMP-dependent Cl- current (I(Cl,cAMP)) leads to cell swelling. We first investigated the mechanism underlying the I(Cl,cAMP)-independent recovery (shrinkage) of the swollen cells. They shrank when the membrane potential (Vm) was made negative to the equilibrium potential of Cl- (ECl) by lowering [K+]o or [Cl-]o in the high-K+ solution. This shrinkage was attenuated by the inhibitors (DIDS, glibenclamide, furosemide) of swelling-activated Cl- current (I(Cl,swell)). These findings suggested an involvement of I(Cl,swell) in the observed isosmotic cell shrinkage. On the other hand, an application of hyposmotic (70% of control) solution to the cells at normal [K+]o (ECl>Vm) induced a cell swelling, and the swollen cells underwent a slight but definite spontaneous cell shrinkage during hyposmotic challenge, indicating the operation of the mechanism of regulatory volume decrease (RVD). This RVD was pronounced at low [Cl-]o, at which ECl was much more positive than Vm. On the contrary, when the hyposmotic solution was applied to the cells at high [K+]o, at which ECl was negative to Vm, the cells swelled vigorously and monotonically without showing RVD, the swelling being much greater than that seen at normal [K+]o. Both the RVD at normal [K+]o and the extra cell swelling at high [K+]o were suppressed by DIDS. These results suggest that I(Cl,swell) activated by cell swelling can shrink or inflate the cardiac cells under hyposmotic as well as isosmotic conditions, depending on Vm and ECl.

Chronic cough is the only symptom of eosinophilic bronchitis (EB). There is considerable overlap between EB and atopic cough. To investigate the antitussive effects of a histamine H1-recetor antagonist, epinastine hydrochloride (epinastine, CAS 80012-43-7, Alesion; 20 mg/day, once daily), cough scores, pulmonary function, capsaicin cough threshold, and bronchial hyperresonsiveness (BHR) to methacholine (MCh) were evaluated before and after a 4-week treatment with epinastine in patients with EB. In the epinastine group, the cough scores were decreased significantly (18.3 +/- 6.1 in week 1, 17.4 +/- 6.7 in week 2, 15.1 +/- 6.2 in week 3, 14.0 +/- 4.8 in week 4) in comparison with the value of 35.3 +/- 8.7 in week -2). The cough threshold for capsaicin improved from 1.70 +/- 3.04 micromol/l to 12.7 +/- 17.6 micromol/l in the epinastine group (p < 0.05; baseline vs. week 4) The bronchial hyperresponsiveness to MCh (Dmin) did not change significantly either in the epinastine or the placebo groups. The morning and evening peak expiratory flow rate (PEFR, L/min) did not change from the baseline period in either the epinastine or the placebo groups. These results suggested that epinastine may be useful for treating patients with EB and that histamine H1-receptor is related to the pathophysiology of coughing in EB.

Fistulojejunostomy was performed at the subcutaneous level in two patients with intractable pancreatic fistula that occurred after surgery for cancer of the ampulla of Vater and carcinoma of the lower bile duct. The treatment yielded mostly satisfactory results, though one patient incurred postoperative wound dehiscence, which was healed with conservative measures. Compared with conventional procedures, this method is technically easy to perform, as it does not involve surgical separation of the fistula up to a site deep within the abdominal cavity, rarely results in side injury, and poses few potential risks of cicatricial stenosis of the fistular lumen, because blood supply to the fistula is preserved. Subcutaneous fistulojejunostomy is considered to be recommendable for pancreatic fistulas that occur long after surgery and which are stenosed at the site of the pancreatojejunal stenosis and thus require surgical treatment.

A sensor for nitric oxide is reported that uses a novel redox matching mechanism to induce a resistance change upon binding this important ligand to cobalt.

Study Objective: Th2 cytokines play an important role in the pathogenesis of asthma. Our study objective was to determine the effect of suplatast tosilate, a Th2 cytokine inhibitor, on patients with cough-variant asthma., Methods: Twenty patients with cough-variant asthma (CVA) were assigned to a suplatast tosilate (100 mg three times daily) group or a placebo group for 6 weeks in a double-blind randomized study. The cough scores, medication scores, pulmonary function, bronchial hyperresponsiveness to methacholine, cough threshold for capsaicin, percentage of eosinophils and concentrations of eosinophilic cationic protein (ECP) in hypertonic saline-induced (induced) sputum were evaluated. The main outcome measures were capsaicin cough threshold and concentrations of ECP in induced sputum., Results: In the suplatast group, the cough scores and the medication scores decreased significantly over time. The percentage of eosinophils in induced sputum significantly decreased from 53.5+/-5.6% to 13.6+/-2.6%. The cough threshold for capsaicin improved significantly from 2.72+/-3.41 microM to 39.7+/-22.7 microM in the suplatast group. The concentrations of ECP in induced sputum decreased significantly from 435+/-123 microg/l to 56+/-34 microg/l in the suplatast group. The bronchial responsiveness to methacholine changed from 8.45+/-3.43 units to 11.4+/-3.76 units in the suplatast group., Conclusions: Suplatast improved the cough scores and the cough threshold for capsaicin in patients with CVA without significant side effects, suggesting the effectiveness of suplatast in the treatment of CVA. Suplatast also decreased the percentage of eosinophils and concentrations of ECP in induced sputum, suggesting improvement in eosinophilic inflammation in patients with CVA. Further pharmacodynamic research is needed to explain the precise mechanism.

Hereditary hemorrhagic telangiectasia (HHT or Rendu-Osler-Weber syndrome) is an autosomal dominant disorder characterized by aberrant vascular development. We report here a genetic epidemiologic study in a county, A, in the Akita prefecture (population 1.2 million) located in northern Japan. Nine HHT patients who had been referred to tertiary-care hospitals were located in and near the study county. A total of 137 pedigree members were traced of which 81 were alive and 32 were affected by HHT. Complications associated with cerebral or pulmonary arteriovenous malformations were proven in six out of seven families. Linkage analysis in two large families revealed a weak yet suggestive linkage to the HHT1 locus (encoding endoglin; ENG). Three novel mutations were found in four families, all of which led to a frameshift: a G to C transversion at the splicing donor site of intron 3 (Inv3+1 G>C) in one family, one base pair insertion (A) at nucleotide 828 (exon 7) of the endoglin cDNA in two large families (c.828-829 ins A), and a four base pair deletion (AAAG) beginning with nucleotide 1120 (exon 8) of the endoglin cDNA (c.1120-1123 delAAAG) in one family. The insertion of A in exon 11 (c.1470-1471 insA) mutation found in one family has also been reported in a European family. No endoglin gene mutations were found in two families. The population prevalence of HHT in the county was estimated to be 1:8,000 approximately 1:5,000, roughly comparable with those reported in European and U.S. populations, which is contradictory to the traditional view that HHT is rare among Asians. We recommend that families with HHT be screened for gene mutations in order that high-risk individuals receive early diagnosis and treatment initiation that will substantially alter their clinical course and prognosis., (Copyright 2002 Wiley-Liss, Inc.)

Carcinomas rarely occur at the site of a colostomy. A 73-year-old man underwent abdominoperineal resection (Miles' surgical procedure) for rectal carcinoma in September 1988. He did not return to the hospital until September 1995, when he was admitted with stricture of the stoma. Neither tumor nor ulcer was detected at the stoma. The tip of the doctor's little finger was able to pass through the stoma, and manual expanding alleviated the stricture. The patient did not return to the hospital again until August 1996, at age 81 years, when he visited the hospital because of complete stricture of the stoma. A biopsy revealed an adenomatous carcinoma, but the results of various examinations suggested no metastasis. A portion of the intestine, including the stoma and surrounding skin, was resected, and a new stoma was created in the descending colon. We report this rare carcinoma occurring at the site of a colostomy, and we review the literature.

In order to clarify the mechanism of the antitussive effects of azelastine hydrochloride (azelastine, CAS 790307-93-0), the cough responses to inhaled capsaicin and substance P (SP) were evaluated before and after the administration of azelastine in conscious guinea pigs. The concentrations of SP were also measured before and after the administration of azelastine by radioimmunoassay in anesthetized guinea pigs. Capsaicin and SP caused coughing in conscious guinea pigs in a dose-dependent fashion. After the treatment with azelastine, capsaicin-induced cough decreased significantly, and the dose-response curve to capsaicin was shifted to a higher concentration in comparison with the the controls. SP-induced cough was not inhibited by the treatment with azelastine, and the dose-response curves to SP did not change. The concentrations of SP recovered in bronchoalveolar lavage fluid and in the trachea were decreased statistically significantly in a dose-dependent manner after the treatment with azelastine, while the SP concentrations of the subjects not treated with azelastine were not inhibited. These results suggest that the antitussive effect of azelastine might be partly due to the inhibition of SP release from sensory nerves in guinea pigs.

Intracellular adhesion molecule-1 (ICAM-1)-mediated cell-cell adhesion is thought to play an important role at sites of inflammation. Recent evidence suggests that ICAM-1 surface expression on alveolar macrophages is increased in pulmonary sarcoidosis and that inflammatory granuloma formation is characterized by the aggregation of macrophages. The present study shows that ICAM-1 expression is significantly elevated on alveolar macrophages from patients with sarcoidosis in response to tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (INF-gamma) compared with healthy controls. Aggregation and adhesion were significantly increased in alveolar macrophages treated with TNF-alpha and INF-gamma, and significantly inhibited in those pretreated with a monoclonal antibody to ICAM-1. Similarly, aggregation and adhesion were inhibited in macrophages treated with heparin, which then exhibited a wide range of biological activities relevant to inflammation. These results suggested that the surface expression of ICAM-1 on alveolar macrophages in response to TNF-alpha and INF-gamma is important in mediating aggregation and adhesion. Additionally, heparin may be useful for developing novel therapeutic agents for fibrotic lung disease.

We examined the role of p38, p42, and p44 mitogen-activated protein kinase (MAPK) isoforms and cytosolic phospholipase A(2) (cPLA(2)) activation in human eosinophil adhesion to plate-coated fibronectin (FN). In the control state, eosinophil adhesion was maximal, with 10 microg/ml FN at 30 min, and decreased after 60-90 min. Western blot analysis demonstrated that p44/42 MAPK (extracellular signal-regulated kinase (ERK)1/2) and cPLA(2) were phosphorylated during adhesion to FN, whereas p38 MAPK phosphorylation was unchanged. Preincubation of eosinophils with U0126 or PD98059, two structurally unrelated MAPK kinase inhibitors, or arachidonic trifluoromethyl ketone, a cPLA(2) inhibitor, blocked eosinophil adhesion to FN. By contrast, eosinophil adhesion was unaffected by SB203580, a p38 MAPK inhibitor. Pretreatment of eosinophils with okadaic acid, a serine/threonine phosphatase inhibitor, at the concentrations that induced ERK1/2 and cPLA(2) phosphorylation caused an increase in maximal eosinophil adhesion to FN for >60 min. MAPK kinase inhibition but not p38 inhibition also blocked FN-mediated F-actin redistribution in eosinophils and prevented cPLA(2) phosphorylation caused by adhesion to FN. These results demonstrate that ERK1/2 mediating cPLA(2) activation is essential for eosinophil adhesion to FN.

We examined the role of a cytosolic phospholipase A2 (cPLA2) in antigen-induced eosinophil infiltration of airways and in airway hyperresponsiveness to methacholine. Inhibition of cPLA2, or blockade of the platelet-activating factor (PAF) receptor, blocked antigen-induced airway hyperresponsiveness and suppressed eosinophil infiltration. Neither cyclooxygenase nor 5-lipoxygenase inhibition had either effect. We show here that, in antigen-sensitized guinea pigs, cPLA2 inhibition prevents both eosinophilic infiltration and subsequent airway hyperresponsiveness after antigen challenge. We also show that this effect is mediated by first-step hydrolysis of membrane phospholipid into lysophospholipid rather than by prostanoid or leukotriene metabolites of arachidonate.

The effects of the activation of cyclic AMP-dependent Cl- current (ICl,cAMP) on cell volume were studied at various [K+]o under isosmotic conditions in guinea-pig ventricular myocytes. The area of the cell image obtained with videomicroscopy was used as an index of cell volume. I(Cl,cAMP) was activated by adrenaline (5.5 microM). Measurements of the membrane potential (Vm) were performed by the gramicidin-perforated patch-clamp method. At 5.4 mM [K+]o with low [Cl-]o, where Vm was negative to the predicted equilibrium potential of Cl- (ECl), adrenaline sizably decreased the cell area. At high [K+]o with normal [Cl-]o, where Vm was positive to ECl, adrenaline increased the cell area; at 145.4 mM [K+]o the cell area was increased to 110% of control on average (n = 22). The cells swollen in this manner shrank when [Cl-]o was reduced to a low level in the presence of adrenaline. The results indicate that the induction of Cl- influxes (outward I(Cl,cAMP)) or effluxes (inward I(Cl,cAMP)) can lead to a cell swelling or shrinkage, respectively. The addition of BaCl2 (1 mm), a blocker of K+ channels, attenuated the adrenaline-dependent cell swelling, supporting the view that Cl- fluxes must be accompanied by cofluxes of K+ ions to affect the cell volume. The adrenaline-dependent cell swelling was inhibited by antagonizing beta-adrenergic stimulation with acetylcholine or by blocking I(Cl,cAMP) channels with glibenclamide, confirming the involvement of I(Cl,cAMP) in the adrenaline response. The results show that the activation of I(Cl,cAMP) can shrink or inflate the cardiac cells under isosmotic conditions, depending on Vm and ECl.

Fibroblast migration, proliferation, extracellular matrix protein synthesis and degradation, all of which play important roles in inflammation, are themselves induced by various growth factors and cytokines. Less is known about the interaction of these substances on lung fibroblast function in pulmonary fibrosis. The goal of this study was to investigate the effects of PDGF alone and in combination with IL-1beta and TNF-alpha on the production of human lung fibroblast matrix metalloproteinases, proliferation, and the chemotactic response. The assay for MMPs activity against FITC labeled type I and IV collagen was based on the specificity of the enzyme cleavage of collagen. Caseinolytis and gelatinolytic activities of secreted proteinases were analyzed by zymography. Fibronectin in conditioned media was measured using human lung fibronectin enzyme immunoassay. Cell proliferation was measured by 3H-Thymidine incorporation assay. Cell culture supernatants were tested for PGE2 content by ELISA. Chemotactic activity was measured using the modified Boyden chamber. Matrix metalloproteinase assay indicated that IL-1beta, TNF-alpha and PDGF induced intestitial collagenase (MMP-1) production. MMP assay also indicated that IL-1beta and TNF-alpha had inhibitory effects on MMP-2,9(gelatinaseA,B) production. Casein zymography confirmed that IL-1beta stimulated stromlysin (matrix metalloproteinase 3; MMP-3) and gelatin zymography demonstrated that TNF-alpha induced MMP-9 production in human lung fibroblast, whereas PDGF alone did not. PDGF in combination with IL-1beta and TNF-alpha induced MMP-3 and MMP-9 activity, as demonstrated by zymography. PDGF stimulated lung fibroblast proliferation in a concentration-dependent manner, whereas IL-1beta and TNF-alpha alone had no effect. In contrast, the proliferation of human lung fibroblasts by PDGF was inhibited in the presence of IL-1beta and TNF-alpha, and this inhibition was not a consequence of any elevation of PGE2. PDGF stimulated fibroblast chemotaxis in a concentration-dependent manner, and this stimulation was augmented by combining PDGF with IL-1beta and TNF-alpha. These findings suggested that PDGF differentially regulated MMPs production in combination with cytokines, and further that MMP assay and zymography had differential sensitivity for detecting MMPs. The presence of cytokines with PDGF appears to modulate the proliferation and chemotaxis of human lung fibroblasts.

Fibroblast migration, proliferation, extracellular matrix protein synthesis and degradation are the key events in various biological and pathological processes in pulmonary fibrosis. In addition, biopsy specimens from the lungs of patients with pulmonary fibrosis show increased numbers of mast cells which have metachromatic granules containing heparin, histamine and proteases. Little is known about how these products influence pulmonary fibrosis. In the present study, we investigated the effect of heparin and related glycosaminoglycans on PDGF-induced lung fibroblast proliferation and chemotactic response in vitro. In addition, we examined the effect of heparin on both the induction of matrix metalloproteinases (MMPs) and MMPs activity in lung fibroblasts in vitro. Heparin, de-N-sulphated heparin but not heparan sulphate inhibited PDGF-induced lung fibroblast proliferation. In contrast, only heparin inhibited PDGF-stimulated human lung fibroblast chemotaxis. Negatively charged poly-L-glutamic acid had no effect on either fibroblast proliferation or chemotaxis. Thus the negative charge alone cannot account for the ant-proliferative and anti-chemotactic effects of heparin. Furthermore, heparin and heparan sulphate also had no inhibitory effect on induction of MMPS, including MMP-1 (interstitial collagenase), MMP-2 (gelatinase A) and MMP-9 (gelatinase B). Only heparin inhibited both MMP-1 and MMP-2/MMP-9 activity. Additionally, tissue inhibitor of metalloproteinase type 1 (TIMP-1) and type 2 (TIMP-2) inhibited PDGF-stimulated human lung fibroblast chemotaxis. The ability of heparin to inhibit fibroblast chemotaxis may account for the inhibitory effect of heparin on MMP activity. The above results suggested that heparin and related glycosaminoglycans differentially regulate PDGF-induced lung fibroblast proliferation, chemotaxis and MMPs activity and further that these effects may have a key role in extracellular matrix remodeling in inflammatory lung disease.

The relationship of 5-FU-sensitivity to thymidylate synthase (TS) was investigated in a total of 82 gastric cancers of stage III or IV. The sensitivity test was done by Histoculture Drug Response Assay and TS expression was stained immunohistochemically using anti-TS antibody. Sensitivity to 5-FU of more than 50% of the inhibitory index (high sensitivity) was observed in 28.1% of the materials, less than 50% (low sensitivity) in 71.9% and less than 20% in 50.0%. The expression of TS was found in 25.6% of the patients. The incidence of positive TS expression was 34.5% in the high sensitivity group, against 22.0% in the low sensitivity group, with no statistical difference between them. No particular relationship was found between 5-FU-sensitivity group and TS expression rate when the tumor-histology was divided into high and low differentiation. In the present study, the ratio of patients negative for TS among the high 5-FU-sensitivity group both was 65.2%, both of which have been reported as producing high survival rates. In contrast, the ratio of patients positive for TS in the low 5-FU-sensitivity group was 22.0%. From these results, TS is considered to be responsible for approximately 40-50% of either high or low 5-FU-sensitivity, respectively, when the lower TS expression in the present study was taken into consideration and corrected.

A 61-year old asymptomatic woman was admitted to our hospital for the examination of an abnormal shadow in the left lower lung lobe in 1978. Enhanced chest computed tomograms and pulmonary arteriograms revealed a pulmonary arteriovenous malformation (PAVM) composed of feeding artery and draining vein. The patient had suffered brain abscesses 3 times because of paradoxical emboli from PAVMs. A diagnosis of hereditary hemorrhagic telangiectasia (HHT) was made according to the criteria. The patient died of septic shock due to urinary tract infection by Candida albicans. We reviewed cases of PAVMs associated with HHT in the Japanese literature. In Japan, 126 HHT families and 144 HHT patients have been reported to date. PAVMs occur in approximately one-third of HHT patients in Japan. Twenty-four out of 45 patients (44.4%) had multiple PAVMs. We also discussed the diagnosis, complications, and treatment of PAVM-associated HHT.

A 17-year-old man was admitted to our hospital because of exertional dyspnea, fever, and hemoptysis. Chest X-ray films disclosed diffuse alveolar infiltrates and patchy shadows in both lungs. Laboratory data included a hemoglobin level of 6.7 g/dl and white blood cell count of 8,100/microliter. Urinalysis revealed microscopic hematuria with RBC cast. Bronchoalveolar lavage fluid from the right middle lobe bronchus was bloody. Anti-GBM antibodies were detected at low levels in serum (10 EU/ml) by ELISA procedures. Renal and lung biopsies were performed. Immunofluorescent studies revealed linear deposits of IgG along glomerular basement membrane of the kidney, but not in alveolar walls of the lung. This case fulfilled the criteria for Goodpasture's syndrome. The patient was treated with methylprednisolone (1,000 mg/day, for 3 days) and plasma exchange (for 2 days), and demonstrated a dramatic improvement in his clinical condition and chest X-ray findings. We were unable to identify autoantibodies to the NC domain of the alpha 3 chain of type IV collagen. Another conformational epitope may play a role in the disease.

We analyzed the binding of heparinoid or heparin with fibrinogen by real-time measurement using surface plasmon resonance technology. Poly(glucosyloxyethyl methacrylate) sulfate [poly(GEMA) sulfate] and dextran sulfate were used as heparinoids. The binding ability of each sulfated polymer was estimated by having each polymer-containing buffer interact with the sensor chip surfaces that had immobilized fibrinogen. Dextran sulfate and poly(GEMA) sulfate showed high affinity to the fibrinogen in this experiment, while the heparin did not. All of the dextran sulfates were desorbed from its surface, while about 30% of the poly(GEMA) sulfate remained on the immobilized fibrinogen upon the addition of NaCl to the buffer which was done in order to analyze the desorption of poly(GEMA) sulfate or dextran sulfate from the surface of the fibrinogen. These data show that the type of binding between fibrinogen-poly(GEMA) sulfate was different from that of dextran sulfate, indicating that the interaction between fibrinogen and poly(GEMA) sulfate was caused not only by an electrostatic but also by a hydrophobic force. These results suggest that the interaction mechanism of heparinoids with fibrinogen was different from that of heparin.

The immunohistochemical expression of thymidylate synthase (TS) and thymidine phosphorylase (TP) was investigated in 116 of early gastric cancer, in order to know whether or not these reflect malignity in an early stage. The materials conditioned on early gastric cancer with submucosal invasion and over 1 cm2 in size, were 57 with and 59 without lymph node metastasis. They were divided into two by the depth of invasion. The expressions of TS and TP in these group were compared with corresponding histopathological findings. Overall expressions of TS and TP were 54.3% and 34.5%, respectively. The TS-expression was not related with the depth of invasion and lymph node metastasis. The TP-expression, however, showed significant difference between with and without lymph node metastasis, and was so on the depth of submucosal invasion in the group without the nodal metastasis. Multivariate analysis showed that mucosal spread bordering 4 cm2 in size (p = 0.024) and lymphatic permeation (p = 0.099) in TS-expression, and lymph node metastasis (p = 0.041), submucosal invasion (p = 0.076) and venous permeation (p = 0.111) in TP-expression were the noticeable factors regarding to their high expression rates. Although these results were considered not to exceed gastric resection on the prognosis, they might be applicable as one of the indicators in postoperative follow-up on the minor resection of early gastric cancer such as EMR or local resection.

Interest in the treatment of the pulmonary arteriovenous malformations (PAVMs) that occur in approximately one-third of patients with Rendu-Osler-Weber (ROW) disease (hereditary haemorrhagic telangiectasia) has recently been renewed. PAVMs can now be occluded safely by the transvenous placement of detachable balloons or metal coils, thus avoiding the many potential complications of thoracotomy. This study analyses the treatment of eight PAVMs in four ROW patients by transcatheter embolization using detachable balloons or metal coils. After embolization, the mean right-to-left shunt fraction significantly decreased from 39.1 +/- 5.1% to 11.9 +/- 1.1% (P < 0.05) and PaO2 significantly increased from 53.3 +/- 7.8 torr to 76.2 +/- 8.4 torr (P < 0.05). No serious complications occurred. One detachable balloon was deflated, but no recanalization occurred. We conclude that transcatheter embolization is a safe and efficacious treatment for PAVMs associated with ROW disease. Long-term studies are now needed to determine the risk of recanalization in this treatment.

Unlabelled: Using a simple technique detecting the eosinophil fraction in whole-blood flow cytometry, we measured intracellular antigen EG2 (a monoclonal antibody to eosinophil cationic protein) in 56 asthmatic patients (26 during an attack and 30 during an asymptomatic period) and 22 healthy subjects to determine whether EG2 reflects the pathological stages of allergy., Methods: In brief, preparations of the sample included the following procedures: (1) hemolyzation of heparinized or EDTA-mixed whole blood; (2) fixation of white blood cells with 0.4% parabenzoquinone (PBQ) or paraformaldehyde (PFA); (3) permeabilizing the cell membrane with n-octyl-beta-D-glucopyranoside, and (4) staining of intracellular EG2 antigen with monoclonal EG2 antibody and FITC-labeled secondary antibody., Results: In PBQ-fixed samples, there was a clearer boundary of the eosinophil fraction with a higher yield and purity than in those fixed with PFA. The number of EG2-positive eosinophils was significantly greater in subjects during attacks than in asymptomatic patients. In addition, when compared with normal controls, asthmatic subjects had significantly greater numbers of EG2-positive eosinophils regardless of their current condition., Conclusion: Eosinophil intracellular EG2 may indicate the pathological stage of asthma. This simple technique for analysing the properties of eosinophils using whole-blood flow cytometry would save time and labor in laboratories.

Reconstruction of the Photon Factory storage ring (PF ring; 2.5 GeV) is now in progress to provide very brilliant synchrotron radiation to users, i.e. the emittance is being reduced by a factor of five. Components, such as the quadrupole and sextupole magnets, vacuum chambers, beamlines and beam-position monitors, are being replaced by new ones in 16 normal-cell sections of the PF ring. The accelerating cavities, injection systems and control systems are also being replaced. Operation will commence when the improvements are completed on 1 October 1997.

The Tristan Main Ring (MR) at KEK (High Energy Accelerator Research Organization) has been used as a third-generation synchrotron radiation ring with an undulator installed in the MR as a third-generation light source. The magnetic field irregularities of the undulator were examined precisely. Measurement of the magnetic field after synchrotron radiation experiments showed that the magnetic irregularities in both the horizontal and vertical directions are negligibly small. This result suggests that our previous estimates of emittance, made using the measured undulator spectrum, should correctly represent the properties of the electron beam in the MR.

To investigate the efficacy of azelastine hydrochloride (azelastine, CAS 79307-93-0, Azeptin) in suppressing cough, 22 bronchial asthma patients complaining mainly of cough were given the drug for four weeks. Peak flow rates (PEFR), pulmonary function tests, capsaicin cough threshold, and bronchial hyperresponsiveness were compared pre- and post-administration. After four-week's administration of azelastine (2 mg twice daily), cough decreased as demonstrated in a significant progressive improvement of cough points. The morning PEFR (1/min) was improved significantly at one week and two weeks post-administration. Changes were from 434 +/- 26.4 pre-administration to 461 +/- 25.8 at Week 1 (p < 0.05), 462 +/- 26.7 at Week 2 (p < 0.05), 452 +/- 22.5 at Week 3, and 462 +/- 20.8 at Week 4. The evening PEFR (1/min) showed 439 +/- 22.2 pre-administration, 454 +/- 21.4 at Week 1, 464 +/- 22.4 at Week 2, 457 +/- 19.3 at Week 3 and 467 +/- 17.8 at Week 4, improvement being significant at Week 1 (p < 0.05). Regarding pulmonary function tests no significant changes were observed. FVC (liter), FEV1 (liter), and FEV1/FVC (%) were 3.45 +/- 0.86, 2.68 +/- 0.52, and 83.6 +/- 5.93 pre-administration; and 3.48 +/- 0.21, 2.72 +/- 0.65, and 84.1 +/- 6.21 post-administration, respectively. The capsaicin cough threshold [Ccap (mumol/l)] showed significant improvement, changing from 5.95 (0.016-50.0) pre-administration to 19.7 (0.08-50.0) post-administration (p < 0.05). Conversely, an index of bronchial hyperresponsiveness, Dmin (mg/dl;U), showed no significant changes (14.9 +/- 5.2 vs. 19.7 +/- 5.3). These results suggest that azelastine inhibits cough in patients with bronchial asthma by increasing the level of the cough threshold without changing bronchial hyperresponsiveness.

The authors evaluated clinical importance of alveolar-arterial PO2 difference (AaDO2) and pulmonary artery pressure (PAP) estimated by pulsed Doppler echocardiography in 31 patients with pulmonary embolism (PE). Echocardiographic estimates from flow velocity patterns in the right ventricular outflow tract showed significant correlation with actual measurements obtained by right cardiac catheterization. Furthermore, PAP as obtained by pulsed echocardiography was significantly higher in acute massive and recurrent multiple groups in comparison with the acute submassive group. AaDO2 was greatest in the acute massive group, followed by the recurrent multiple group, and then by the acute submassive group. These results suggest that analyses of AaDO2 and the echocardiographic estimation of PAP at bedside are successful in the diagnosis and classification of PE.

Background: It is believed that the incidence of cystic fibrosis (CF) among Asiatic races, including the Japanese, is very rare. This epidemiological study was carried out to investigate the incidence of CF in Japan., Methods: We collected literature describing CF cases among pure Japanese and found 124 cases reported as CF during the 43 years from 1951, when the first case was reported, to 1993. Only 104 cases (57 male and 47 female patients) of 124 cases met our diagnostic criteria., Results: A simple calculation based on the number of reported CF cases and of live births after 1980 suggested that the incidence of CF is about 1 in 350,000 in the Japanese population. Twenty-nine (27.9% of the total) of 30 patients diagnosed in the neonatal period presented symptoms of meconium ileus, an incidence higher than that reported for the white population., Conclusions: Our study results suggest that the incidence of CF in the Japanese population is even rarer than had been estimated before and that there is a genetic difference between northern European and Japanese populations.

A 5.4 m-long 239-pole undulator comprising three rigid and precise standardized-unit undulators was completed and installed in the Tristan main ring, which was operated as a light source at a beam energy from 8 to 10 GeV. This undulator was successfully tested, characterized and used as a brilliant hard X-ray source for several synchrotron radiation experiments during the last 3.5 months of 1995.

A 35-year-old woman was referred to our hospital for investigation of lower abdominal pain and a feeling of fullness. At her first consultation, a transvaginal ultrasonography (US) revealed a homogeneous cystic mass in the lower abdomen. Over a period of 8 months the US findings of the content of this mass changed from fine and faint internal echoes to moderate amounts of irregularly contoured internal echoes. At laparotomy, the cystic mass, which measured 3.0 x 3.0 x 3.5 cm, appeared to arise in the sigmoid mesocolon and tightly adhered to the appendix. The cyst was unilocular and contained a slightly yellow gelatinous fluid. Microscopically, its wall was fibrous and lacked an epithelial lining, suggesting that it was a so-called pseudocyst arising from the sigmoid mesocolon. To our knowledge, this is the first case report documenting the time-course ultrasonographic observations of a mesenteric pseudocyst. Our findings suggest that the time-elapsed ultrasonographic changes might have been dependent on the interval between the onset of cystic formation and the US examination.

The short-term and long-term effects of the anticholinergic agent oxitropium bromide (CAS 30286-75-0, OTB) were studied in nine elderly patients with bronchial asthma (8 men and 1 women, age: 69.5 +/- 5.2 years, 3 atopic and 6 (not atopic cases). 2 h after inhalation of 0.8 mg of OTB, forced vital capacity (FVC) had increased from 2.28 +/- 0.22 1 to 2.87 +/- 0.27 1 (p < 0.01), forced expiratory volume in 1 s (FEV1) had increased from 1.22 +/- 0.14 to 1.57 +/- 0.16 1 (p < 0.01), and respiratory resistance (Rrs) had decreased from 5.9 +/- 0.8 cmH2O/l/s to 4.9 +/- 0.7 cmH2O/l/s (p < 0.05). The patients inhaled OTB (0.2 mg t.i.d.) for 6 weeks. Their peak expiratory flow rates, measured each morning and each evening, increased significantly during the first 2 weeks of administration, and the increase was sustained for the entire weeks (p < 0.05). Other pulmonary-function tests were done before drug administration and again at the end of the 6 weeks. By the end of the 6th week of OTB administration, FVC had increased from 2.30 +/- 0.22 1 to 2.99 +/- 0.26 1 (p < 0.01), and FEV1 had increased from 1.25 +/- 0.17 to 1.70 +/- 0.22 1 (p < 0.05). These data indicate that OTB can be useful in the treatment of elderly patients with bronchial asthma.

To assess the effects of azelastine in patients with cough-variant asthma, we measured the cough threshold for capsaicin (the concentration required to elicit more than five coughs) in 16 patients with cough-variant asthma before and after 4 weeks of treatment with azelastine (2 mg; b.i.d.) or placebo. After treatment, coughing decreased in all patients and the cough threshold for capsaicin increased significantly, from 0.67 +/- 0.30 microM to 4.76 +/- 1.55 microM (P < 0.01) in the azelastine group. However, the cough threshold for capsaicin did not increase significantly, from 0.86 +/- 0.33 microM to 1.11 +/- 0.35 microM (P > 0.10) in the placebo group. These results suggest that azelastine inhibits coughing in patients with cough-variant asthma.

The effect of FK224 (N-(N2-[N-¿N-(N-2,3-didehydro-N-methyl-N-[N-3- (2-pentylphenyl)-propionyl¿-L-threonyl]tyrosyl-L-leucynyl)-D -phenylalanyl¿-L-allothreonyl]-L-asparaginyl)-L-serine-v-lacto ne, CAS 125787-94-2) on isometric contraction of canine tracheal smooth muscle in situ was studied. Contraction was induced by administration of substance P, neurokinin A, and neurokinin B intra-arterially into the tracheal circulation in five mongrel dogs. FK224 inhibited substance P- and neurokinin A-induced contraction in a dose-dependent manner, but it did not inhibit neurokinin B-induced contraction significantly. These data suggest that FK224 is a dual antagonist of both neurokinin 1 and neurokinin 2 receptors, with a similar potency in in vivo experiments.

Two autopsy cases (69 and 87-year-old women) of malignant mixed müllerian tumor (MMMT) following radiation therapy for uterine cervical cancer sixteen and twenty years ago respectively were reported. They were admitted due to abdominal pain and diagnosed as ileus. In the first case, CT examination revealed a tumor measuring about 8 x 10 cm in size in the uterine posterior wall. Recurrence of the uterine cancer was suspected and hysterooophorectomy combined with sigmoidectomy was performed. In the second case, artificial anus formation was performed because of sigmoid stricture by the invasion of the tumor. Histologically, tumors in both cases were composed of carcinomatous and sarcomatous components including heterologous elements such as cartilage. The patients died of extensive spreads and metastasis in the liver and alimentary tract.

The efficiency of the 13C-Urea Breath Test (13C-UBT) for the detection of Helicobacter pylori colonization in gastric mucosa was evaluated. The 13C-UBT was performed in five pediatric and six adult subjects who had had upper gastrointestinal endoscopy within 2 weeks. H. pylori colonization was confirmed in two pediatric and three adult subjects with peptic ulcer combined with antral gastritis, by histological examination of antral biopsy specimens. When an individual with H. pylori colonization ingested a solution containing 13C-urea, a significant amount of 13CO2 appeared in the respiratory CO2 within 10 min. The mean cumulative percentage dose of 13C recovered in the breath over 30 min in the cases with H. pylori colonization was significantly higher than that in those who were not colonized (4.91 vs 0.41, P < 0.001). In addition, the effect of antibiotic on the eradication of H. pylori from gastric mucosa was monitored by 13C-UBT in two cases. The values of cumulative percentage dose of 13C over 30 min fell to the same levels as those observed in H. pylori negative subjects after just 2 weeks treatment with amoxicillin; however, positive results were obtained again 1 month after the withdrawal of amoxicillin. In summary, 13C-UBT is a simple, reliable, non-invasive method in the diagnosis of gastric H. pylori colonization especially for pediatric patients.

Quantitative analysis and immunohistochemical studies of small intestinal mucosa were performed to investigate the mechanism of mucosal damage in 10 patients with food-sensitive enteropathy. Jejunal biopsy specimens were taken before and after treatment and after clinical relapse following a challenge test. The low villous height of untreated patients normalized after introduction of an elimination diet but declined again to subnormal level after a challenge test. Several other types of cells were significantly increased in the untreated patients in comparison to controls. These included HLA-DR+ (DR+) CD4+ cells in the lamina propria and intraepithelial CD8+ cells. Moreover, those cell patterns, such as increased DR+ CD4+ cells and CD8+ cells, normalized with treatment but regressed to pretreatment levels when the patients were challenged. These findings suggest that activated CD4+ cells in the lamina propria of the small intestinal mucosa, probably by releasing cytokines, may play an important role in contributing to mucosal damage in patients with food-sensitive enteropathy.

To evaluate the role of Helicobacter pylori in the pathogenesis of gastritis and peptic ulcer in children, ultrastructural changes of the gastric mucosa with H. pylori colonization were studied in two pediatric patients with duodenal ulcers. The study demonstrated that H. pylori: (i) colonized the mucous layer covering normal gastric epithelium; (ii) lay atop short, irregular microvilli and at the intercellular junctions of damaged cells; and (iii) adhered firmly by specific junction zone (firm connection) and loosely by filamentous appendages to the epithelial surfaces of cells which lacked evidence of a protective mucous layer. The ultrastructural changes, which become progressively worse from patterns (i) to (iii) mentioned above, suggest that these patterns represent distinct and successive stages of the infection. These findings, therefore, lend support to the notion that H. pylori infection is quite common and may relate to the pathogenesis of gastritis and peptic ulcer in children as well as in adults.

An 18-year-old male with a lifelong history of atopic dermatitis and recurrent severe Staphylococcus aureus abscesses was found to have hyperimmunoglobulinemia E. We measured IFN-gamma production by peripheral blood mononuclear cells cultured with a protein A non-producing strain of staphylococcal antigen and serum specific IgG subclass antibodies to this antigen in this patient. IFN-gamma production, when cultured with non protein A produced by this antigen in this patient was almost the same as that in healthy subjects. IFN gamma m-RNA was detected at the dose of 0.0625 micrograms total RNA. The levels of IgG1, IgG3 and IgG4 were higher in this patient than in healthy subjects, while IgG2 was lower. We thus identified a case of hyperimmunoglobulinemia E syndrome with normal IFN-gamma production.

We have studied the effect of neurokinin A (NKA) on tracheal smooth muscle (TSM) contraction caused by administration of acetylcholine (ACh) intra-arterially (ia) into the tracheal circulation, or bilateral stimulation of the vagus nerves, in 26 mongrel dogs. The tracheal contractile response to 10(-7) mol NKA was 3.12 +/- 0.48 g/cm, 6.3 +/- 0.7%AChmax. Plasma histamine concentration was measured in the right atrium at the peak response following administration of NKA. Plasma histamine concentration was 3.33 +/- 2.05 ng/ml after 10(-7) mol NKA (vs. 1.82 +/- 0.54 ng/ml for control, P > 0.10). A frequency-response curve was generated by electrical stimulation of the caudal ends of cut cervical vagi over the range of frequencies 1-20 Hz (constant 30 V) at 15 s intervals 10 min before and 10 min after administration of 10(-8) mol NKA. Substantial augmentation of tracheal contractile responses to 10(-10)-10(-7) mol ia ACh was obtained 10 min after administration of NKA (P < 0.05). Significant potentiation of tracheal contraction to vagal stimulation for 10-20 Hz was also observed after administration of 10(-8) mol NKA (P < 0.05). We have demonstrated a substantial contractile effect of NKA on canine tracheal smooth muscle that is not related to histamine release from respiratory mast cells. We have also demonstrated that NKA causes augmentation of the parasympathetic tracheal contractile response induced by ia administration of ACh and efferent vagus nerve stimulation. These data suggest that potentiation of vagal contractile response by NKA is related to post-synaptic activation of the parasympathetic nerves.

The present patch-clamp study shows that external Mg2+, Ca2+ and Sr2+ decrease the unit amplitude of inward current through the inward-rectifier K+ channel in a concentration-dependent manner. Sr2+ produces a voltage-dependent flickering block as well, and the fractional electrical distance between the external orifice and the Sr2+ binding site (delta) is 0.73. The decrease of unit amplitude is reversible and voltage independent while it does not increase the noise level on the open-channel current. Unit current decreased by Mg2+ or Ca2+ has a longer mean open time, which is inversely proportional to the unit amplitude. External Mg2+ does not decrease the amplitude of unit outward current. A surface potential shift, measured using voltage-dependent Cs+ block (delta = 1.60), failed to explain the current decrease. Therefore, we conclude that (1) the external divalent cations cause an extremely fast channel block, which appears as a decreased amplitude of the unit current on the recording system; (2) the blocking site (fast site) is present near the external orifice of the channel, and it is separate from the blocking site (slow site) to which Cs+ and Sr2+ bind.

A 53-year-old mentally handicapped female with hereditary hemorrhagic telangiectasia complicated by pulmonary arteriovenous fistula developed a recurrent brain abscess in the left frontal region. She responded well to surgical drainage and antibiotic therapy. Previously, she had two brain abscesses of the left parieto-occipital and right parietal regions removed on separate occasions at 38 and 40 years of age. Although pulmonary arteriovenous fistula was eventually confirmed by angiography at age 41 years, surgical removal of the pulmonary lesion was not indicated due to multiplicity and large vascular shunting. Early recognition of pulmonary arteriovenous fistula in patients with hereditary hemorrhagic telangiectasia is emphasized for diagnosis of brain abscess and prevention of recurrence.

We studied the effect of neurokinin A (NKA) on tracheal smooth muscle (TSM) contraction induced by the administration of acetylcholine (ACh) intra-arterially (ia) into the tracheal circulation or bilateral vagal stimulation in 18 mongrel dogs. In five dogs, 10(-12) to 10(-7) mol of NKA was injected selectively into the tracheal circulation. The tracheal contractile response to 10(-7) mol NKA was 3.12 +/- 0.48 g/cm, 6.3 +/- 0.7% AChmax. Plasma histamine concentration in the right atrium was measured at the peak response to NKA. Plasma histamine concentration was 3.33 +/- 2.05 ng/ml after 10(-7) mol NKA (vs 0.32 +/- 0.08 ng/ml for control, p > 0.10). In the other 5 dogs, dose-response was obtained with 10(-12) to 10(-7) mol ACh 10 min before and 10 min after administration of 10(-8) mol NKA. Frequency-response relationship also was induced by electrical stimulation of the distal cut ends of the cervical vagi in the range of frequencies 1-20 Hz (constant 30 V) at 15-s intervals 10 min before and after administration of 10(-8) NKA. Considerable potentiation of the tracheal contractile response to 10(-18) to 10(-7) mol ia ACh occurred 10 min after administration of 10(-8) mol NKA (p < 0.05). Significant potentiation of the tracheal contraction to vagal stimulation at 10-20 Hz was also observed 10 min after administration of 10(-8) mol NKA (p < 0.05). We demonstrate that the substantial contractile effects of NKA on canine tracheal smooth muscle are not related to histamine release from mast cells in the respiratory tract.(ABSTRACT TRUNCATED AT 250 WORDS)

In order to evaluate isometric tension in segments of tracheal smooth muscle in vivo, we first investigated the anatomical distribution of arteries supplying the tracheal smooth muscle in 52 dogs. The patterns of the cranial thyroid arteries were classified into the following four types. A) Common type: the cranial thyroid artery leaves the muscular branch, and then divides into the thyroid branch and the tracheal branch. B) The cranial thyroid artery leaves the tracheal branch, and then divides into the muscular branch and the thyroid branch. C) The cranial thyroid artery divides into the muscular branch, the thyroid branch, and the tracheal branch. D) The cranial thyroid artery divides into the tracheal branch and thyroid branch, and lacks a muscular branch. Out of 52 dogs, 62% had common type bilaterally, and 23% had common type unilaterally. All dogs had a tracheal branch. We demonstrated tracheal smooth muscle contraction by intra-arterial administration of acetylcholine, histamine, 5-hydroxy-tryptamine, phenylephrine, and clonidine using isometric technique in vivo. These results suggest that tracheal contraction induced by sympathetic amines is mediated through two subtypes of receptors, alpha 1 and alpha 2, in tracheal smooth muscle.

The purpose of this study was to clarify the bronchodilating effect of pirenzepine (PZ) and to verify its mechanism. Ten asthmatic patients (6 men, 4 women: aged 20 to 65, 5 atopic 5 non-atopic) and ten non-asthmatic volunteers (8 men, 2 women: aged 25 to 60) were studied. Forced vital capacity (FVC), forced expiratory volume in one second (FEV1.0) and peak expiratory flow rate (PEFR) were measured after intravenous administration of 20 mg PZ. PZ increased FVC, FEV1.0 and PEFR significantly by 15%, 29% and 37% respectively in asthmatic patients (p less than 0.01). We also studied the effects of PZ on the contractile responses of tracheal smooth muscle to intra-arterially administered acetylcholine (ACh) and the electrical stimulation of the vagus nerves (VNS) using isometric technique in situ in 5 mongrel dogs. PZ significantly inhibited the contractile responses elicited with ACh at doses larger than 1000 micrograms/kg (p less than 0.01). PZ also significantly inhibited the contractile responses elicited by VNS at doses larger than 100 micrograms/kg (p less than 0.01). These data demonstrate that intravenously administered PZ dilates the airway in asthmatic patients and also suggest that the bronchodilating effect of PZ related to inhibition of the M1 and M3 muscarinic receptors.

This study was undertaken to investigate cytokine production of mononuclear cells (MNCs) from patients with bronchial asthma stimulated by antigens of Candida albicans in vitro. The reaction of MNCs from corticosteroid-dependent patients was restricted to a low level. The level of tumor necrosis factor (TNF)-alpha released into the culture fluid was significantly higher in the high responders (HR) of the atopic corticosteroid-independent (A-CSID) group than those of other asthmatic and control groups (p less than 0.01). The level of IL-1 beta of the A-CSID group was significantly higher than that of the non-atopic corticosteroid-independent group (p less than 0.05), but not significantly different from the controls. In HR of the A-CSID group, the production of TNF-alpha and IL-1 beta was augmented and interferon-gamma production was also increased in these patients. These results suggest that Candida albicans can contribute to the augmentation of cytokine production in bronchial asthma, especially in some of atopic type.