Poly I:C-induced expression of intercellular adhesion molecule-1 in intestinal epithelial cells.

Intercellular adhesion molecul-1 (ICAM-1) is a transmembrane glycoprotein belonging to the immunoglobulin superfamily of adhesion molecules and plays perdominant roles in recruitment and trafficking of leucocytes to sites of inflammation. ICAM-1 expression in intestinal epithelial cells (IECs) is enhanced by several stimuli, such as proinflammatory cytokines, bacterial infections or pathogen-associated molecular patterns. One of these stimuli, double-stranded RNA (dsRNA), is a by-product of viral replication and can be recognized by its cognate receptor Toll-like receptor 3 (TLR-3). In spite of expression of both TLR-3 and ICAM-1 in IECs, correlation between TLR-3-signalling and ICAM-1 expression has never been examined in IECs. In the present study, we investigated whether poly I:C, an analogue of dsRNA, can stimulate the expression of ICAM-1 in IEC line, HT-29. Poly I:C-stimulation up-regulated the expression of ICAM-1 mRNA by real-time polymerase chain reaction. Enhanced expression of ICAM-1 was confirmed in protein level by immunofluoresense cell staining and enzyme-linked immunosorbent assay by measuring the released soluble ICAM-1 in culture supernatant. As the stimulation effect was reduced by pre-treatment of the cells with anti-TLR-3 antibody, poly I:C-binding signal was thought to be sensed by TLR-3 on the surface of HT-29. The results of luciferase assay and nuclear factor kappa-b (NF-kappaB) inhibitor treatment experiments indicated that the downstream signal was mainly transduced by transcription factor, NF-kappaB. All these results demonstrated the connection between TLR-3 signalling and ICAM-1 expression in HT-29 cells and indicated the importance of coordinated function of both innate and adaptive immunity against viral infections.

Characterization of human dental pulp-derived cell lines.

AIM: To establish and characterize different types of fibroblastic cell lines derived from dental pulp tissue. METHODOLOGY: Human dental pulp tissue-derived cells were transfected with SV40 large T antigen by Lipofectamine transfection method. Geneticin (G418)-resistant cells were selected and different cell lines were established by a limiting dilution method. To characterize the lineages of cells, each clone was immunofluorescently stained by anti-fibroblast, anti-vimentin, anti-collagen type I and type III antibodies. Total RNA was extracted from each clone and subjected to a differential display experiment. RESULTS: By transfecting SV40 large T antigen, nine different cell clones were obtained. All these cell clones were positively stained by anti-fibroblast, anti-vimentin, anti-collagen type I and type III antibodies. With differential display experiment, eight different genes, the expression levels of these genes were varied amongst each cell clone, were detected. After sequencing and database search, one gene was revealed to be identical to T-cell marker, Thy-1. Thy-1 expression in dental pulp tissue was confirmed by immunohistochemical staining. CONCLUSION: Fibroblastic cell lines derived from human dental pulp tissue possessed different gene expression profiles suggesting the existence of subpopulations.

Active synthesis of mouse polymeric immunoglobulin receptor in the epithelial cells of the distal urinary tubule in kidney.

The tissue distribution of mouse polymeric immunoglobulin receptor (pIgR) has been demonstrated. By Northern blot hybridization, pIgR mRNA expression was detected in liver, intestine, stomach, lung and kidney. A weak expression was also detected in thymus by reverse transcriptase-polymerase chain reaction. The pIgR expression in kidney was further studied and confirmed that pIgR protein was actively synthesized in the epithelial cells of distal urinary tubule and of Henle's loop. Immunoelectron microscopical analysis showed the accumulation of pIgR-containing vesicles in the apical portion of distal urinary tubule epithelial cells.

[Effect of a leukotriene antagonist, pranlukast hydrate, on exercise-induced asthma for 14 days].

Exercise is a common stimulus of bronchoconstriction in subjects with asthma. Exercise-induced asthma (EIA) may be caused by heat loss and/or water loss during exercise. These changes cause the release of mediators and the development of bronchoconstriction. Recent studies have implicated the cysteinyl leukotrienes in the pathogenesis of EIA. Therefore we investigated the effect of a leukotriene antagonist, pranlukast hydrate, on EIA. Subjects were 11 EIA positive patients and were all atopic type bronchial asthma. 450 mg of pranlukast hydrate was given for 14 days and the exercise test was performed before and after treatment. Pulmonary function tests were done before and 0, 5, 15 and 30 min after completion of the each exercise. Prostaglandin E2, thromboxane B2, leukotriene E4 and B4 in urine were measured before and after the treatment. Pranlukast hydrate did not affect FEV1.0, PF and Flow volume curve at rest and also levels of mediators in urine. The mean maximal percentage fall in FEV1.0 after exercise was--54.9 +/- 5.7% without therapy and--30.1 +/- 7.5% after pranlukast hydrate therapy. There was a significant difference between FEV1.0 values before and treatment at each time (p < 0.05). This study demonstrated that pretreatment with a potent and selective leukotriene antagonist, pranlukast hydrate, attenuated EIA, and it suggests that leukotriene plays a role in EIA.

[Effects of eicosapentaenoic acid in patients with bronchial asthma].

Eicosapentaenoic acid (EPA) is composed of 20 unsaturated fatty acids and is similar to arachidonic acid. Epadel, the drug made from EPA, is reported not only to reduce levels of serum lipids, but also to have antiinflammatory and antiallergic effects. EPA may exert these effects via control of the production of prostanoids and leukotrienes. We studied the effects of EPA in patients with bronchial asthma who had hyperlipidemia. The patents were given EPA at 1800 mg/day and they recorded signs and symptoms in an asthma diary during a 2-week observation period and the 8 weeks during which they took the drug. Peak flow, leukotriene B4 concentration in urine, Leukotriene E4 concentration in urine, IgE level, total cholesterol level, and triglyceride level were measured before and after the 8 weeks of medication. Administration of EPA was associated with improvements in symptom score, therapeutic score, asthma score, and peak flow. EPA may be useful in patients with asthma complicated by hyperlipidemia.

[Immunologic analysis of milk, hen egg, and soybean proteins in butter and margarine, and clinical assessment for availability of hypoallergenic margarine (HAM)].

To determine the allergenic proteins in commercially available butter and margarine, protein fractions were extracted and immunologically analyzed against milk, hen egg, and soybean antigens. Butter and 10 kinds of margarine were proved to contain these proteins in various concentrations by ELISA and immunoblotting methods by use of the rabbit antisera developed against each food proteins. However, hypoallergenic margarine was found to contain no such proteins at all. Using sera obtained from atopic dermatitis patients, previously detected high levels of IgE antibodies to milk, hen egg, or soybeans, the extracted protein fraction from butter and margarine was analyzed whether these proteins react with IgG antibodies in patients' sera. The sera with high levels of specific IgE recognized protein antigens in these extracts except those from hypoallergenic margarine, suggesting that proteins in butter and margarine may become allergenic for the patients sensitive to the proteins, and that hypoallergenic margarine will be a reasonable alternative for the allergic patients to milk, hen egg, or soybeans.

[Effects of a beta 2-agonist, sodium cromoglycate, and an anticholinergic agent on hyperventilation-induced bronchoconstriction in sensitized rabbits].

Hyperventilation can induce bronchoconstriction in ovalbumin-sensitized rabbits. To investigate the roles of the beta-receptor parasympathetic nervous system and of chemical mediators in hyperventilation-induced bronchoconstriction (HIB), the effects of a beta 2-agonist, of sodium cromoglycate, and of an anticholinergic agent on HIB were studied. Rabbits were divided to four groups and treated as follows. Group 1: Control (n = 7, 0.9% saline); Group 2: Procaterol (n = 4, 50 micrograms/l); Group 3: Sodium cromoglycate (n = 4, 10 mg/ml); and Group 4: Ipratropium bromide (n = 6, 1 mg/ml). Each drug was inhaled for 1 min via an ultrasonic nebulizer. Then, for the eucapnic hyperventilation challenge, sensitized rabbits were mechanically hyperventilated for 15 min (120 breaths/min, tidal volume = 7 ml/kg) with dry air containing 5% CO2 at room temperature. Total lung resistance and dynamic compliance were measured before, and 0, 5, 15, and 30 min after hyperventilation. The mean percent change in resistance measured 5 min after the hyperventilation was +49% in group 1, -6% in group 2, +23% in group 3, and +1% in group 4. The changes in groups 2 and 4 were significantly less than in group 1 (p < 0.05). In conclusion, HIB is mainly caused by bronchial smooth muscle constriction, and chemical mediators and the parasympathetic nervous system may play important roles in the development of HIB in sensitized rabbits.

[Effect of combination therapy with disodium cromoglycate (DSCG) and Ozagrel on non-atopic asthmatics].

Ozagrel (OKY-046), a selective inhibitor of thromboxane biosynthesis has been reported to reduce airway hyperresponsiveness in asthmatics. DSCG has been widely used for atopic, but not for non-atopic asthmatics. We evaluated the additive effect of combination therapy with DSCG and Ozagrel on nonatopic asthmatics. Fourteen asthmatics were divided into two groups. The patients in the O-D group were initially treated with Ozagrel (400 mg/day) for 4 weeks, and then DSCG (8 mg/day) was added for another 4 weeks. The patients in D-O group were initially treated with DSCG (8 mg/day), and then Ozagrel (400 mg/day) was added for 4 more weeks. TXB2, PGF2 alpha, PGE2, 6-keto-PGF1 alpha in sputum and 11-dehydro TXB2 in urine were measured before and after 4th and 8th weeks of therapy. Ozagrel decreased the metabolites of thromboxane and PGF2 alpha in sputum and urine. PGE2 in sputum tended to be increased by Ozagrel. DSCG did not affect mediators in sputum. Combination therapy with Ozagrel and DSCG showed the additive effect to single use on attack score and PF in non-atopic asthmatics.

Elevated levels of soluble ICAM-1 in sera from patients with bronchial asthma.

We measured the levels of soluble intercellular adhesion molecule-1 (sICAM-1) in sera from patients with bronchial asthma. sICAM-1 levels in sera from atopic asthmatic patients in stable conditions were higher than in normal control subjects. Furthermore, the sICAM-1 levels in sera obtained during bronchial asthma attacks were higher than those in sera obtained in stable conditions. These results suggest that higher levels of sICAM-1 in sera reflect the upregulation of ICAM-1 expression in allergic inflammation.

[A case of allergic bronchopulmonary aspergillosis with no history of bronchial asthma].

A 53-year-old male was admitted to our hospital because of an abnormal shadow in the left upper lung. Bronchofiberscopy revealed edematous mucosa and pus at the orifice of the left upper lobe bronchus. Pathological examination revealed bronchial inflammatory change with infiltration of eosinophils and also the existence of aspergillus in the pus. Mild eosinophilia and elevation of serum IgE level were observed in the peripheral blood, and serum precipitin against Aspergillus fumigatus was positive. Bronchogram showed central bronchiectasis, and the diagnosis of allergic bronchopulmonary aspergillosis (ABPA) was made. In cases of ABPA, bronchial asthma is usually present prior to presentation, but this patient had no history of asthma even though airway hyperresponsiveness to methacholine was confirmed.

Mass trial of hypoallergenic rice (HRS-1) produced by enzymatic digestion in atopic dermatitis with suspected rice allergy. HRS-1 Research Group.

The clinical usefulness of a hypoallergenic rice (HRS-1) which was produced by enzymatic decomposition of the constituent proteins of original rice was evaluated in a multicentre study in 44 subjects with recalcitrant atopic dermatitis (AD), who were suspected of having rice allergy. The subjects were fed for at least 4 weeks with HRS-1 instead of eliminating both regular rice and wheat from their daily diet. The extent of overall skin lesions was expressed by using the AD affected area and severity index (ADASI). A statistically significant decrease in ADASI was observed at the 2nd and the 4th week readings and at the end of the study. A provocation test with regular rice was carried out in 5 of 44 subjects following the HRS-1 therapy. An obvious increase in ADASI was found in all of these 5 cases just after this procedure. On final evaluation, 77% of the patients tested showed 'moderate' to 'remarkable' improvement, and 59% of the patients a 'moderate' to 'remarkable' reduction in use of the steroid ointment concomitantly used for the treatment. Finally, HRS-1 was evaluated as 'useful' or 'very useful' in 69% of the subjects.

[Clinical effect of hypoallergenic rice (HRS-1) in atopic dermatitis. HRS-1 Research Group].

The usefulness of hypoallergenic rice (HRS-1) was clinically evaluated in 43 patients with severe atopic dermatitis (AD), who were suspected of having rice allergy, in collaboration with 13 hospitals. The patients were fed with HRS-1 instead of eliminating both regular rice and wheat from their daily diet. AD area and severity index (ADASI) was calculated as an indicator of the degree of cutaneous symptoms. Significant decrease of ADASI were observed in the 2nd and 4th week and at the end of the replacement therapy (5.6 weeks on average). On final evaluation, 74% of the patients tested showed "moderate" to "remarkable" improvement, and in 53% of the patients HRS-1 resulted in a "moderate" to "remarkable" reduction in the dosage and the grade of potency of the steroid ointment concomitantly used for the treatment. Finally, HRS-1 was evaluated as "useful" to "very useful" as the elimination diet in 70% of the patients.

[Successful high-dose methylprednisolone therapy in a patient with primary myelofibrosis].

We are presenting a patient with primary myelofibrosis who responded to the High-Dose methylprednisolone therapy (1 g/day for 3 days). Three and a half years ago, a 55-year-old male was admitted to our hospital because of severe erythroblastic anemia, thrombocytopenia, splenomegaly and "dry tap" of bone marrow aspiration. Bone marrow biopsy revealed a marked fibrosis without any blastoid cell proliferations. Since a thrombocytopenia was progressive and refractory to the ordinary therapy, high-dose methylprednisolone therapy was performed which was followed by an administration of activated vitamin D3. After the therapy, hematologic improvements were achieved within a month (RBC: 284 x 10(4)/microliters----413 x 10(4)/microliters, WBC: 3,000/microliters----11,500/microliters, Plat.: 7,000/microliters----20,000/microliters). Three months after the therapy, the bone marrow biopsy and 113In scintigraphy were performed. These tests also proved marked improvement of histological features of the bone marrow and a decrease of uptake of 113In to the spleen, respectively. The patient continues to be in good condition and he is free from any medications at present time.

A method for the determination of tritium in environmental water samples by electrolytic enrichment.

In order to determine the tritium concentration in environmental water samples, the electrolytic enrichment was carried out with (St) and without (S) addition of tritiated water of a certain concentration (deuterium-free) to the samples. With the use of the fundamental formulas on electrolytic enrichment, the deuterium concentration (Dit) before electrolysis for an environmental water sample is determined by liquid scintillation counting and densitometry for the sample St. Furthermore, the tritium concentration in the environmental water sample is determined by the above methods for the sample S, and by the substitution of Dit for Di in the formulas. Tritium concentrations in environmental water samples were found to be determined within an accuracy of 10% by this method when Vi/Vf was 14-25. It is considered that this method dispenses with the direct measurement of low deuterium concentrations (Di) before electrolysis, a special technique on the purification of water for densitometry, and moreover, excludes the possibility of cross contamination in the electrolytic enrichment by the spike cell method.