House dust mite sublingual tablet is effective and safe in patients with allergic rhinitis.

BACKGROUND: House dust mite (HDM) is the major indoor allergen for allergic diseases such as allergic rhinitis (AR) and asthma. Although sublingual immunotherapy is a curative treatment for HDM-induced AR, data from large-scale studies are limited. We evaluated the efficacy and safety of HDM tablets in adolescent and adult patients (aged 12-64 years) with HDM-induced AR with or without intermittent asthma. METHODS: In a double-blind trial in Japan, 968 subjects were randomized 1 : 1 : 1 to 300 index of reactivity (IR), 500 IR, or placebo groups. The primary endpoint was the Average Adjusted Symptom Score (AASS) in the last eight weeks of the 52-week treatment. Secondary endpoints included individual nasal and ocular symptom scores, rescue medication use, and the Japanese Rhinoconjunctivitis Quality of Life Questionnaire (JRQLQ) scores. RESULTS: The AASS in the last eight weeks of treatment significantly improved in both the 300 IR and the 500 IR groups compared to that in the placebo group (P < 0.001). In the 300 IR group, the onset of action occurred at week 8-10. All four nasal symptoms significantly improved in both active treatment groups; rescue medication use and JRQLQ outcome improved in the 300 IR group. Most adverse events (AEs) were mild, and 16 serious AEs (SAEs) were reported; however, none of them were drug-related. CONCLUSIONS: One-year treatment with 300 IR and 500 IR HDM tablets was effective without major safety concerns. The recommended therapeutic dose for AR is 300 IR.

Variants in the 17q21 asthma susceptibility locus are associated with allergic rhinitis in the Japanese population.

BACKGROUND: Allergic rhinitis (AR) is a very common disorder peaking in the teenage years that is mediated by hypersensitivity responses to environmental allergens. Although it is well established that the ORMDL3 locus at chromosome 17q21 is associated with susceptibility to bronchial asthma, the genetic influences of the polymorphisms of the locus in allergic rhinitis are unclear. OBJECTIVE: To examine whether the polymorphisms in the 17q21 asthma susceptibility locus are associated with allergic rhinitis in the Japanese population. METHODS: We performed linkage disequilibrium (LD) mapping of the locus using the HapMap database and conducted an association study of the locus with a total of 15 tag SNPs in two independent populations. We further evaluated correlations of genotypes with changes in expression of genes at the region in lymphoblastoid cell lines in the Japanese population and assessed the expression levels of the genes in nasal epithelium and various human tissues. RESULTS: We found a significant association between a total of five polymorphisms in the 17q21 asthma susceptibility locus, rs9303277, rs7216389, rs7224129, rs3744246, and rs4794820, and AR (minimum P(combined)  = 0.00074, rs4794820). The expression level of the ORMDL3 transcript was significantly correlated with the genotype of rs12150079, rs7216389, rs3744246, and rs4794820 with P < 0.01 (minimum P = 0.0058, rs7216389), and ORMDL3 mRNA was highly expressed in nasal epithelium. CONCLUSION: Genetic variants in the 17q21 asthma susceptibility locus are significantly associated with AR in the Japanese population.

Drug delivery observation of hydrophobe ferrofluid and magnetite nanoparticals by SPring-8 synchrotron radiation.

In this study, the composite magnetic nanoparticles of coated SiO nano film with about 8 nm size and high saturation magnetization value, were synthesized by liquid phase precipitation method. The magnetic nanoparticles can be dispersed in various liquid media, widely known as magnetic fluids or ferrofluids with both magnetic and liquid properties. The materials been collected great interests and more and more attentions to focus into Drug Delivery System (DDS) as a new technology in this paper. We use the composite nanoparticles to disperse H2O and inject the solutions into rat's in-vivo organs. And, in the experiments by using a strong photon beam of SPring-8 Synchrotron Radiation facility, the distribution stat and the effects of magnetic field as well as drug delivery behaviour of nanoparticles in the rat' kidney are verified by the in-vivo observations.

Role of macrophage-stimulating protein and its receptor, RON tyrosine kinase, in ciliary motility.

Macrophage-stimulating protein (MSP) is an 80-kD serum protein with homology to hepatocyte growth factor (HGF). Its receptor, RON tyrosine kinase, is a new member of the HGF receptor family. The MSP-RON signaling pathway has been implicated in the functional regulation of mononuclear phagocytes. However, the function of this pathway in other types of cells has not been elucidated. Here we show that in contrast to the HGF receptor, which was expressed at the basolateral surface, RON was localized at the apical surface of ciliated epithelia in the airways and oviduct. In addition, MSP was found in the bronchoalveolar space at biologically significant concentrations. MSP bound to RON on normal human bronchial epithelial cells with a high affinity (Kd = 0.5 nM) and induced autophosphorylation of RON. Activation of RON by MSP led to a significant increase in ciliary beat frequency of human nasal cilia. These findings indicate that the ciliated epithelium of the mucociliary transport apparatus is a novel target of MSP. Ciliary motility is critical for mucociliary transport. Our findings suggest that the MSP-RON signaling pathway is a novel regulatory system of mucociliary function and might be involved in the host defense and fertilization.

Inhibition of experimental pulmonary metastasis in mice by beta-cyclodextrin-benzaldehyde.

The effect of beta-cyclodextrin-benzaldehyde (CDBA) on experimental pulmonary metastasis in C3H/He mice was examined. In an in vitro assay, the growth of RCT(+) cells was inhibited by 1200 micrograms/ml CDBA using unrenewed media, and by 600 micrograms/ml CDBA in that using daily renewed media. When mice were treated daily with CDBA, 3 weeks later the number of lung nodules developing after i.v. injection of 1 X 10(6) RCT(+) cells was significantly decreased in a dose-dependent manner, i.e., 73.8%, 85.6%, and 95.7% inhibition was observed following 0.5, 5, and 25 mg CDBA/mouse per day p.o. administration, respectively. The same mice showed almost as much natural killer (NK) activity as normal mice. Therefore, experiments were designed to evaluate the effect of CDBA on the NK activity of tumor-free mice whose immunity had been suppressed by 5-fluorouracil (5FU). Injections of 5FU only suppressed this activity to about 50% of normal mice, but the combined treatment with CDBA negated the suppressive effect of 5FU on NK activity. The results suggested that the inhibition of experimental pulmonary metastasis might be induced by the possible combined effects of CDBA; that is, the direct inhibition of tumors and the augmentation of NK cell activity.

Relation of H-2 expression on murine RCT(+) sarcoma cells to lung colonization and sensitivity to NK cells.

Murine RCT(+) sarcoma cells were sorted using a fluorescence-activated cell sorter with regard to the expression of H-2 antigens and then an increased H-2-expressing subclone was established, and named RCT(+)H-2+. The experimental metastasis of RCT(+) cells was compared with that of RCT(+)H-2+ cells by counting pulmonary colonies on the 21st day after i.v. inoculation of tumor cells (5-10 x 10(4)/mouse). When mice were inoculated with RCT(+) cells, mean numbers of pulmonary colonies were 2.1(range 0-6), 2.8(range 0-7) using 5 x 10(4) and 1 x 10(5) cells, respectively. On the other hand, in the mice inoculated with RCT(+)H-2+ cells, figures obtained were 7.0(range 4-16), 31.9(range 13-79), using 5 x 10(4) and 1 x 10(5) cells, respectively. The survival rate of RCT(+)H-2+ cells was higher than that of RCT(+) cells, when this was assayed in the early stage after i.v. injection of 51Cr-labeled cells (1 x 10(5) cells/mouse). In addition, RCT(+)H-2+ cells were more resistant than RCT(+) cells to lysis mediated by natural killer cells. These data suggest that an increase in metastatic ability was paralleled by an increase in the H-2 antigen expression and a decrease in sensitivity to the natural killer cells.

Histamine application to the nasal mucosa induces release of calcitonin gene-related peptide and substance P from peripheral terminals of trigeminal ganglion: a morphological study in the guinea pig.

Short-term effects of application of histamine to the nasal mucosa on trigeminal ganglion neurons containing calcitonin gene-related peptide (CGRP) and substance P (SP) were examined in guinea pig. Immunoreactivities to CGRP and SP in these neurons were decreased 30 min after the histamine application. The decreases were most marked at 1-3 h after application, after which the immunoreactivities began to increase, reaching the base line by 6 h after the application. The immunoreactivities to CGRP and SP in the nerve endings of nasal mucosa were not decreased. The expression of mRNAs for both peptides in the soma of trigeminal neurons was unchanged. The histamine application to the nasal mucosa may cause release of CGRP and SP from terminals of peripheral processes of trigeminal ganglion neurons, and enhance axonal transport of these peptides, but does not affect their biosynthesis in the soma of trigeminal ganglion neurons.

Dynamic helical CT of T1 and T2 glottic carcinomas: predictive value for local control with radiation therapy.

BACKGROUND AND PURPOSE: Tumor volume and cartilage invasion have been suggested as prognostic factors of glottic carcinomas following definitive radiation therapy. Radiologic examinations provide additional information regarding the deep extension of tumor. We determined whether dynamic helical CT can predict local control of early (T1 and T2 stage) glottic carcinomas treated with definitive radiation therapy. METHODS: Sixty-eight patients with early glottic carcinoma evaluated on pretreatment dynamic helical CT were treated with definitive radiation therapy. Tumor detectability, maximum dimension, tumor volume, and involvement of anatomic subsites (anterior commissure, ventricle, subglottic region, and thyroid and arytenoid cartilages) were determined by consensus by three radiologists without previous knowledge of the clinical information. The CT findings were correlated with local control. RESULTS: The two-year local control rate was 76%; 91% for T1 and 60% for T2 lesions. Univariate analysis revealed clinical T stage, tumor detectability, maximum dimension, tumor volume, anterior commissure involvement, ventricle involvement, and thyroid cartilage involvement as significant prognostic factors. Thyroid cartilage involvement was an independent predictor by multivariate analysis. The lesions separate from the thyroid cartilage had a 95% probability of local control, whereas the lesions adjacent to the cartilage had only a 42% control rate. CONCLUSION: Dynamic helical CT provides prognostic information for the results of definitive radiation therapy. Patients with a tumor adjacent to the thyroid cartilage had an increased risk of local failure.

Effects of PAF on histamine H1 receptor mRNA expression in rat trigeminal ganglia.

The application of platelet-activating factor (PAF) to the nasal mucosa of humans has been shown to increase histamine-induced hyper-reactivity. To test the hypothesis that PAF acts by increasing the reactivity of sensory nerve endings in the nasal mucosa to histamine, we examined PAF-stimulated rat trigeminal nerve ganglion cells. We found that relatively low concentrations of PAF (10(-12)-10(-9) M) induced increased histamine H1 receptor mRNA expression. This increase appeared as early as 1 h after PAF stimulation, peaked at 4 h, and disappeared after 24 h. The PAF receptor antagonist WEB2086 inhibited the increased expression of histamine H1 receptor mRNA induced by PAF, suggesting that the effects of PAF are mediated by specific receptors. This PAF effect was abolished by actinomycin D, suggesting that PAF induces de novo transcription of histamine H1 and/or PAF receptor mRNA. PAF may be important in the hyper-responsiveness of nasal mucosa exposed to histamine.

PAF- and histamine-receptor antagonists lessen allergen-induced hearing impairment in guinea pigs.

We have demonstrated degranulation of mast cells in the endolymphatic sac as well as an increase in audiological threshold shift in the experimental animal models following antigen provocation. Mast cells, however, release such chemical mediators as histamine, platelet activating factor (PAF), and leukotriene due to an antigen-antibody reaction on the cell surface. The aim of this study was to clarify the major chemical mediators responsible for hearing impairment in the animal models following antigen provocation. Guinea pigs were actively sensitized with DNP-Ascaris and provoked with an injection of DNP-BSA. A significant audiological threshold shift was observed at 1, 10, 24, and 72 h following challenge with allergen. The peak shift was at 10 h; all changes were reversed after 7 days. This threshold shift was abolished by prior injection of either a histamine- or PAF-receptor antagonist to allergen, but not of a leukotriene-receptor antagonist. Results suggest that histamine and PAF are involved in the hearing impairment induced by allergen exposure in the guinea pig.

Expression of genes MAGE-1, -2, and -3 by human maxillary carcinoma cells.

We analysed the expression of melanoma antigen-encoding (MAGE) gene-1, -2, and -3 in 20 maxillary carcinomas consisting of two cell lines: freshly isolated cancer cells from specimens from 13 patients, and 5 biopsy specimens. The cells were subjected to reverse transcription by the polymerase chain reaction. Fourteen (70%) out of 20 maxillary carcinomas expressed at least one of the MAGE genes. In contrast, five control samples of inflammed mucosa from the maxillary sinus of patients with chronic sinusitis were all negative for the expression of these genes. Results indicated that patients with maxillary carcinoma may be good candidates for specific immunotherapy.

Low doses of anticancer drugs increase susceptibility of tumor cells to lysis by autologous killer cells.

Pretreatment of squamous cell carcinoma (SCC) cells from four patients with low doses of cisplatin, carboplatin or 5-fluorouracil increased the susceptibility to lysis by autologous killer cells in vitro. Exposure of two SCC cell lines to low doses of these drugs increased the cell surface expression of both HLA class I and intercellular adhesion molecule-1 (ICAM-1). HLA class II, neural cell adhesion molecule and B7 were not expressed on the cell surface before or after such treatment. The results suggest that these drugs increase the susceptibility of tumor cells to autologous cell-mediated cytotoxicity, at least in part, by enhancing the expression of HLA class I and ICAM-1.

Maintenance of the differentiated type II cell characteristics by culture on an acellular human amnion membrane.

We have developed a Culture system for guinea pig alveolar type II cells using an epithelium-denuded human amnion membrane as a substratum. The differentiated morphology was maintained for 3 wk by both air-interface feeding and immersion feeding when type II cells were cultured on the basement membrane side of the amnion with fibroblasts on the opposite side (coculture). Functionally high levels of surfactant protein B (SP-B) and C (SP-C) messenger ribonucleic acids (mRNAs) were expressed even after the 3-wk cultivation and surfactant protein A mRNA was detected on day 10 of the culture. The differentiation was also maintained when fibroblasts were cultured on lower chambers of the culture plates (separate culture). In contrast, culture of type II cells without fibroblasts (monoculture) could not preserve the mature morphology. When the monoculture was supplemented with keratinocyte growth factor or hepatocyte growth factor, a monolayer of rather cuboidal type II cells with apical microvilli was maintained. However, the percent area of lamellar bodies in these cells was significantly less than that in freshly isolated type II cells, and mRNA expressions of SP-B and SP-C were also considerably suppressed. These findings suggest that other growth factors or combinations of these factors are necessary for the maintenance of the differentiated phenotype. As substratum, a permeable collagen membrane or a thin gel layer of Engelbreth-Holm-Swarm mouse sarcoma extracts did not preserve the mature characteristics. This culture system using an acellular human amnion membrane may provide novel models for research in type II cells.

Application of organic solvent-soluble membrane filters in the preconcentration and determination of trace elements: spectrophotometric determination of phosphorus as phosphomolybdenum blue.

A simple and rapid preconcentration technique, based on collecting trace elements on a membrane filter and dissolving the membrane filter in an organic solvent, has been applied to the spectrophotometric determination of phosphorus in water. Phosphorus, 0.5-7 mug in 50-500 ml of water sample, is collected as phosphomolybdenum blue on a nitrocellulose or acetylcellulose membrane in the presence of n-dodecyltrimethylammonium bromide, the membrane is dissolved in 5 ml of dimethyl-sulphoxide (DMSO), and the absorbance of the DMSO solution is measured at 710 nm against a reagent blank. Moderate concentrations of silicate, anionic and non-ionic surfactants and high concentrations of sodium chloride do not interfere. Interference from arsenate can be eliminated by reducing the arsenate to arsenite. Condensed and organic phosphates can be determined if they are first converted into orthophosphoric acid by digestion with persulphate. The limit of determination is 0.002 mug of phosphorus in 100 ml of sample.

Audiological study in guinea pigs with type I allergy induced in the inner ear.

Time course studies of electrocochleography and the auditory brain stem response were performed in guinea pigs that were passively sensitized by sera containing antidinitrophenyl reaginic antibody and specifically challenged by dinitrophenyl-bovine serum albumin injected through the stylomastoid foramen. A negative summating potential on electrocochleography was observed from 12 to 48 hours, but not at 72 hours, after the specific challenge. A threshold increase on the auditory brain stem response was observed 15 minutes after the specific challenge; the threshold recovered to the prechallenge level within 7 days. Further, we used Tranilast, a blocking agent of chemical mediator release from mast cells, before the specific challenge. A negative summating potential and head deviation were not observed after the use of this agent. These results suggest that the auditory change provoked in the inner ear of the sensitized guinea pig may have been induced by type I allergy.

Adoptive immunotherapy for head and neck cancer with killer cells induced by stimulation with autologous or allogeneic tumour cells and recombinant interleukin-2.

Peripheral blood lymphocytes drawn by leukapheresis using Haemonetics V50 were mixed and cultured with autologous or allogeneic tumour cell line to activate killer cells by tumour antigenic stimulation, and further with recombinant interleukin-2 (rIL-2). Killer cells were intra-arterially infused, as a primary therapy, in 5 patients with maxillary and one with lingual cancer (squamous cell carcinoma). Effects on reduction of primary tumour size were significantly high without any severe side effects. The effects were interpreted mainly by direct day-by-day observation of the site, findings of CT and histology. Histological findings of the tissue obtained by surgical operation performed after adoptive immunotherapy were remarkable changes, such as infiltration of lymphoid cells around the cancer nets, degeneration of cancer cells, infiltration of scavenger macrophages (giant cells) and so on. The results suggested that adoptive immunotherapy by the killer cells can be a powerful treatment to bring the cancer under control, in with combination of other therapies.

Expression of the MAGE-1, -2, -3, -4, and -6 genes in non-squamous cell carcinoma lesions of the head and neck.

The messenger RNA level of several MAGE genes, some of which have been proven to encode tumor rejection antigens recognized by cytotoxic T lymphocytes, were examined in 41 benign and malignant lesions of the head and neck region. By a reverse transcription-polymerase chain reaction assay and Southern blot hybridization, MAGE-1, -2, -3, -4, and -6 genes were expressed in 25%, 41.7%, 33.3%, 8.3% and 33.3% of 12 non-squamous cell carcinomas, respectively. These tumors consisted of 6 papillary adenocarcinomas, 3 adenoid cystic carcinomas, 2 adenocarcinomas, and 1 mucoepidermoid tumor. Of 7 non-Hodgkin's lymphomas, one case from the oropharynx and 2 from the nasopharynx expressed for the MAGE-1 and MAGE-2 genes, respectively. In contrast, none of 12 benign tumors expressed any of these MAGE genes. Interestingly, of 10 other lesions including hyperplasia, keratosis, and ulcer, one histologically diagnosed as dysplasia expressed the MAGE-2, -3, -4, and -6 genes. These results suggest that the MAGE genes may be expressed in malignant tumors and precancerous lesions but not in benign tumors. In addition, non-squamous cell carcinomas may be suitable targets for specific immunotherapy against MAGE gene products.

Muscarinic acetylcholine receptors on human lymphocytes in patients with Menière's disease.

To investigate patients with Menière's disease and the association of cholinergic hyperreactivity, we performed muscarinic acetylcholine receptor assay using peripheral blood lymphocytes from patients with Menière's disease and non-dizzy, non-allergic control subjects. Cholinergic receptor maximal bindings (Bmax) and dissociation constants (Kd) were compared between the two groups, indicating the number and the affinities of the receptors, respectively. The receptor Bmax value in Meniére's patients during the remission state (108.6 +/- 51.2 fmol/l x 10(6) lymphocytes) was higher than that in normal controls (45.8 +/- 9.2 fmol/l x 10(6) lymphocytes) (p < 0.01). Furthermore, during an exacerbated state, Bmax was increased significantly (223.7 +/- 90.2 fmol/l x 10(6) lymphocytes) compared to the remission state (p < 0.01). In contrast, Kd values for the receptor did not differ between the two groups. These results suggest that patients with Menière's disease have cholinergic hyperreactivity, which may be further upregulated during a state of exacerbation due to an increase in the number of cholinergic receptors.

Type I allergy in the inner ear of the guinea pig.

Guinea pigs were passively sensitized by sera containing antidinitrophenyl reaginic antibody and specifically challenged by dinitrophenyl-bovine serum albumin injected through the stylomastoid foramen. Nystagmus, head deviation, negative summating potentials on electrocochleography, and an increase of threshold and wave I peak latency on auditory brain stem response testing were observed after local challenge. These physiologic changes were reversible and resolved within several days. We also used Tranilast before the specific challenge. It is a blocking agent of chemical mediator release from mast cells. Negative summating potentials and head deviation were not observed after the use of this agent. In the animals that showed physiologic changes, we observed endolymphatic hydrops, mast cell degranulation, and eosinophil infiltration histologically in the challenged side of the inner ear. These results suggest that the physiologic and histologic changes provoked in the inner ear of the sensitized animals may have been induced by type I allergy.

Nasal tissue eosinophils in allergic rhinitis.

The density characteristics and functional heterogeneity of nasal tissue eosinophils were studied. The density distribution profiles of eosinophils from patients with allergic rhinitis (AR) showed peaks at densities of 1.068 to 1.084 g/ml, significantly lower than the densities of eosinophils in non-allergic patients with nasal polyps and chronic sinusitis (p < 0.01). The proportion of hypodense eosinophils in patients with AR was 43%; this was significantly greater than that in non-allergic subjects (p < 0.001). Patients with AR tended to have more EG2-positive tissue eosinophils. Furthermore, normodense eosinophils in nasal tissue tended to show a higher percentage of EG2-positive cells than hypodense eosinophils. On the other hand, circulating hypodense eosinophils showed a higher percentage of EG2-positive cells than normal density eosinophils. These results suggest that tissue eosinophils may be activated, and that the functional heterogeneity of eosinophils is dependent on factors other than cell density.