Atmospheric Humidity Influences Oviposition Rate of Tetranychus urticae (Acari: Tetranychidae) Through Morphological Responses of Host Cucumis sativus Leaves.

We investigated the effects of morphology of host cucumber, Cucumis sativus L., leaves acclimatized to different atmospheric humidity levels on oviposition by adult females of the twospotted spider mite, Tetranychus urticae Koch. Cucumber seedlings were grown at a vapor pressure deficit (VPD) of 0.4, 1.9, or 3.0 kPa at 28°C (90%, 50%, or 20% relative humidity, respectively) in growth chambers until the second true leaves had expanded. Adult females of T. urticae were released on the adaxial surfaces of leaf squares cut from first and second true leaves in each treatment group, and held in the same humidity condition. Eggs were counted 2 d after release. The lower acclimatization humidity (higher VPD) increased trichome (leaf hair) density of the host leaves and oviposition rate, but the relationship between the trichome and oviposition differed between leaf positions. The leaf mass per area (LMA) was greater in first true leaves than in second true leaves, but was not influenced by VPD. A linear regression model with oviposition rate as the dependent variable and trichome density and LMA as independent variables showed that both variables influenced the oviposition rate approximately equally. We conclude that oviposition was accelerated under low humidity (high VPD) conditions indirectly probably through an increase in the trichome density of host leaves.

Fucoidan derived from Cladosiphon okamuranus Tokida ameliorates murine chronic colitis through the down-regulation of interleukin-6 production on colonic epithelial cells.

Our previous study indicated that the interleukin (IL)-6/STAT-3 signal was up-regulated in inflammatory bowel disease (IBD) in both humans and animal models. We also discovered phosphorylated STAT-3 in the nucleus of the colonic epithelial cells in IBD mice. Intestinal epithelial cells (IEC) have been shown to secrete IL-6. Therefore, the secretion of IL-6 from IEC may be one of the mechanisms of STAT-3 phosphorylation in IEC during the pathogenesis of IBD, and inhibition of IL-6 production by IEC may be beneficial in preventing IBD. We examined the preventative effect of various types of fucoidans on IL-6 production in a lipopolysaccharide (LPS)-stimulated murine colonic epithelial cells line, CMT-93, in vitro. We also determined in vivo the effect of fucoidans on murine chronic colitis induced with dextran sodium sulphate. Among fucoidans, those from Cladosiphon okamuranus Tokida and Kjellmaniella crassifolia inhibited IL-6 production in CMT-93 cells with the down-regulation of NF-kappaB nuclear translocation. Analysis of the effect of fucoidan on murine colitis in vivo showed that the disease activity index and myeloperoxidase activity decreased in mice fed Cladosiphon fucoidan, but not Fucus fucoidan. Cytokine profiles in colonic lamina propria indicated that the synthesis of interferon (IFN)-gamma and IL-6 decreased and that of IL-10 and transforming growth factor (TGF)-beta increased in mice fed Cladosiphon fucoidan, compared with mice fed a standard diet or Fucus fucoidan. The levels of IL-6 mRNA in colonic epithelial cells was lower in colitis-induced Balb/c mice fed Cladosiphon fucoidan than those fed a standard diet. Fucoidan improves murine chronic colitis by down-regulating the synthesis of IL-6 in the colonic epithelial cells. Fucoidan derived from C. o. Tokida may be useful as a dietary substance for the patients with inflammatory bowel disease.

Water monitoring system for oil contamination using polymer-coated quartz crystal microbalance chemical sensor.

A water monitoring system with a new chemical sensor for oil contamination was developed. The sensor had an organic polymer film on a quartz crystal microbalance (QCM). The organic film was a hydrocarbon polymer and had high affinities for the organic compounds of petroleum products such as gasoline, kerosene, diesel oil and fuel oil. The monitoring system as composed of a sampling part, a purging part, a humidity control part and a sensor part. The oil in contaminated river water could be detected whose threshold odour number (TON) was less than three. The detecting time was less than 5 min depending on the oil kindness. This system was tested using artificially contaminated river water with the oils, to be found that the sensitivity was kept steady for longer than 6 months with 400 detections of diesel oil and heavy oil. Moreover, the oil kind could be discriminated with only one sensor device by analyzing the desorption response curves obtained by flowing a clean air on the sensor instead of the purging air.

Warthin's tumor of parotid gland on Tc-99m pertechnetate scintigraphy with lemon juice stimulation: Tc-99m uptake, size, and pathologic correlation.

The aim of this study was to evaluate the usefulness of technetium-99m (Tc-99m) pertechnetate scintigraphy with lemon juice stimulation in the diagnosis of Warthin's tumor and its correlation with Tc-99m uptake, tumor size, and histologic subtype. Tc-99m pertechnetate scintigraphy before and after lemon juice stimulation and pathologic specimens of 34 Warthin's tumors and 47 non-Warthin's lesions were retrospectively evaluated. Tc-99m uptake of Warthin's tumors before and after stimulation was visually graded as follows: absent; indeterminate; low grade; definite; and strong. Tumor size was defined as maximum diameter of the tumor measured from the surgical specimen. Warthin's tumors were classified into three histologic subtypes according to the ratio of epithelial and lymphoid stromal components: predominant epithelial; intermediate; and low-grade epithelial types. Eighteen of 34 (53%) Warthin's tumors and one benign lymphoepithelial cyst showed higher uptake than that of the normal parotid gland on Tc-99m scintigraphy before lemon juice stimulation. Thirty-two of the 34 (94%) Warthin's tumors, one benign lymphoepithelial cyst, one pleomorphic adenoma, and one oncocytoma revealed higher uptake than that of the normal parotid gland on Tc-99m scintigraphy after lemon juice stimulation. The mean size was 37 mm in strong uptake Warthin's tumors, 24 mm in definite uptake tumors, 19 mm in low-grade uptake tumors, and 12 mm in low-grade uptake tumors excluding those tumors with large cystic component. There was a significant correlation between tumor size and degree of Tc-99m uptake after lemon juice stimulation. However, there was no correlation between histologic subtype and Tc-99m uptake, and histologic subtype and tumor size in Warthin's tumors. Our study concludes that Tc-99m pertechnetate scintigraphy with lemon juice stimulation is useful for the detection and diagnosis of Warthin's tumor. The degree of uptake in Warthin's tumor on Tc-99m scintigraphy with lemon juice stimulation depends mainly on tumor size and the presence of large cystic component in it.

Mucosal immunity of the middle ear: analysis at the single cell level.

OBJECTIVE: Studies have suggested that the middle ear is a potential site of immunological regulation and that the middle ear mucosa constitutes a part of the mucosal immune system. We clarify the characteristics of the middle ear mucosa with respect to immune potential. STUDY DESIGN: We investigated lymphocyte subsets, mRNA of cytokines, and induction of antigen-specific IgA-producing cells in the middle ear mucosa in specific pathogen-free C57BL/6 mice. RESULTS: Flow cytometric analysis showed a certain amount (10%-15%) of gammadelta T cells among CD3+ T cells. P6-specific IgA-producing cells were induced by intranasal immunization with P6 together with cholera toxin. RT-PCR assay of mucosal T cells detected mRNA of Th2 type cytokines such as IL-5 and IL-10. CONCLUSION: These findings support the fact that the middle ear is potentially an effector site of the mucosal immunity.

Immortalization of rat middle ear epithelial cells by adeno 12-SV40 hybrid virus.

Rat middle ear epithelial cells were infected with the adeno 12-SV40 hybrid virus. The cell line thus obtained displays features of primary cultured epithelial cells in both light microscopic and ultrastructural examinations. The immortalized cells have been in continuous proliferation for 40 passages and more than 17 months. Immunohistochemical analysis of the immortalized cells was positive for the SV40 T antigen and the tumor suppressor protein p53. The cells also stained positive for cytokeratin, an epithelial cell marker, and negative for vimentin, a fibroblast marker. These results, together with karyotype analysis, indicate that this cell line originated from rat middle ear epithelial cells and retains the characteristics of epithelial cells. This cell line will be useful for studying the normal cellular biology of middle ear epithelial cells, as well as the cellular and molecular mechanisms involved in the bacteria-middle ear epithelial cell interaction.

Properties of fucoidan from Cladosiphon okamuranus tokida in gastric mucosal protection.

To elucidate the anti-ulcer potential of Cladosiphon fucoidan, anti-peptic activity, bFGF stabilizing activity and inflammatory properties of this and related substances were investigated. Anti-peptic activity was observed with this and other sulfated polysaccharides such as dextran sulfate, carrageenan, and Fucus fucoidan. However, non-sulfated polysaccharides such as mannan and dextran did not exert the anti-peptic activity. The loss of bFGF bioactivity was prevented by all sulfated polysaccharides tested except chondroitin sulfate, at pH 7.4 and at pH 4.0. At pH 2.0, only heparin protected the bFGF activity. The generation of superoxide by macrophages and PMNs was stimulated by dextran sulfate, carrageenan, and Fucus fucoidan, whereas Cladosiphon fucoidan, heparin and chondroitin did not. Dextran sulfate, carrageenan, and Fucus fucoidan also stimulated the secretion of TNFalpha from macrophages, while Cladosiphon fucoidan did not. Thus, Cladosiphon fucoidan is a sulfated polysaccharide without inflammatory action. These results suggest that Cladosiphon fucoidan is a safe substance with potential for gastric protection.

Structural study of fucoidan from Cladosiphon okamuranus TOKIDA.

A structural study was carried out on a fucoidan isolated from the brown seaweed Cladosiphon okamuranus. The polysaccharide contained fucose, glucuronic acid and sulfate in a molar ratio of about 6.1 : 1.0 : 2.9. The results of Smith degradation showed that this polysaccharide has a linear backbone of 1-->3-linked alpha-fucopyranose with a half sulfate substitution at the 4-positions, and a portion of the fucose residues was O-acetylated. The data obtained from partial acid hydrolysis, a methylation analysis and NMR spectra indicated that the alpha-glucuronic acid residue is linked to the 2-positions of the fucose residues, which were not substituted by a sulfate group. These results indicated that the average structure of this fucoidan is as follows: -[(-->3Fuc-4(+/-OSO3-)alpha1-)5-->3[GlcA alpha1-->2]Fuc alpha1-]n-. (Half of each fucose residue was sulfated. One O-acetyl ester was present in every 6 fucose residues.)

Inhibitory effect of Cladosiphon fucoidan on the adhesion of Helicobacter pylori to human gastric cells.

We studied the inhibitory effect of Cladosiphon fucoidan on the attachment of Helicobacter pylori (H. pylori), a gastroduodenal pathogen, to human gastric cell lines. The bacterial binding in these cell lines was inhibited more by Cladosiphon fucoidan (IC50 = 16-30 mg/mL), than by the fucoidan from Fucus (IC50 > 30 mg/mL). Dextran sulfate, another sulfated polysaccharide, did not inhibit the binding at all. Pre-incubating the bacterial suspension with fucoidans reinforced the inhibitory ability of these components, and reduced the IC50 value of Cladosiphon fucoidan to approximately 1 mg/mL. However, the binding was not inhibited by pre-treatment of gastric cells with these components. It was also shown that this fucoidan blocks both Leb- and sulfatide-mediated attachment of H. pylori to gastric cells. Furthermore, fucoidan-binding proteins were found on the H. pylori cell surface by Western blot analysis. Thus, the inhibitory effect exerted by Cladosiphon fucoidan on binding between H. pylori and gastric cells might result from the coating with this component of the bacterial surface.

The role of immune complex in otitis media with effusion.

Fifteen chinchillas were inoculated with immune complexes of Streptococcus pneumoniae into the tympanic cavity, and with formalin-killed bacteria into the opposite side. Middle ear effusion (MEE) were found in 14 ears having inoculation with immune complexes and in only four ears having inoculation with formalin-killed S. pneumoniae. In another experiment, 20 chinchillas received intra-tympanic inoculation with live S. pneumoniae after systemic immunization with formalin-killed S. pneumoniae of the same strain. As a control, 12 chinchillas were inoculated with live bacteria of the same strain into the tympanic cavity without pre-immunization. MEE was observed in nine of pre-immunized animals and persistent MEE lasting for more than 3 weeks was observed in seven of the animals. In control, persistent MEE was not observed. Though six animals developed otitis media with effusion (OME). The results suggest that the formation of immune complexes in the tympanic cavity plays an important role in the occurrence of persistent MEE after pneumococcal otitis media.

Gardner syndrome in a boy with interstitial deletion of the long arm of chromosome 5.

We described a 15-year-old boy with Gardner syndrome (GS), mental retardation, and craniofacial abnormalities. High-resolution banding analysis showed an interstitial deletion of the long arm of chromosome 5 (q22.1----q31.1). The breakpoints in the present case and in 3 previously reported 5q- patients with adenomatous polyposis coli suggest that the gene responsible for GS/or familial polyposis coli (FPC) is in the 5q22 region, a result consistent with the findings of linkage studies.

[The removal efficiency of beta 2-microglobulin, alpha 1-microglobulin and alpha 1-acid glycoprotein by using dialyzers in the bloodstream and transference of endotoxin through membranes].

Retention of beta 2-microglobulin (beta 2-MG) has been indicated as one of the causes of hemodialysis-associated amyloidosis. Membranes with higher permeability (high-performance membrane) have recently been developed accordingly. Several dialyzers were tested by us in an attempt to study their removal efficiency of beta 2-MG and higher molecular substances of alpha 1-microglobulin (alpha 1-MG) and alpha 1-acid glycoprotein (alpha 1-AG), as well as transferring of endotoxin (ET) through membranes into bloodstream. The membranes subjected to our study were PMMA, CTA, PS, EVAL-C and CA. Removal efficiency of beta 2-MG through EVAL-C and CA was lower as compared with that through other membranes (P less than 0.05), while the EVAL-C showed relatively higher removal efficiency for alpha 1-MG and alpha 1-AG. Permeability for ET was not observed with all membranes studied herein.

Influence of middle ear effusion on perilymph.

To investigate the influence of middle ear effusion (MEE) on perilymph (PL), an experimental otitis media with effusion (OME) was provoked in chinchillas by injecting the tympanic cavity with immune complexes. MEE remained for up to 9 days after the injection of immune complexes. PL was aspirated on the 4th, 10th, and 21st day after the inoculation. The mean concentrations of albumin, IgG, and prostaglandin E2 (PGE2) were significantly greater in PL from OME induced ears than in that form normal control ears. 3H-PGE2 placed on the round window membrane of pathologically affected ears passed into PL in significantly greater amounts than in normal controls. The findings of the present study indicate that MEE affects the biochemical environment of PL. However, this influence was transient and reversible.

Experimental otitis media in mice. Immunohistochemical observations.

In order to identify T cells and B cells in the middle ear (ME) mucosa, immunohistochemical observations were performed on mice which were induced with immune-mediated otitis media with effusion (OME). C3H/HeN mice were challenged with dinitrophenylated ovalbumin in the ME, 2 weeks after the systemic immunization. At 1, 3 and 7 days post challenge, the mice were sacrificed, and IgG-, IgA- or IgM-positive cells, as well as Lyt-1- or Lyt-2-positive cells, were observed in the subepithelial layer of ME. IgG- or IgA-positive lymphocytes were detected at 7 days post challenge, whereas IgM-positive cells were rarely seen. At 3 days post challenge, the number of Lyt-1-positive lymphocytes exceeded that of Lyt-2-positive lymphocytes; however, at 7 days post challenge they were similar. Previous studies have suggested that immune complexes are a key factor in the generation of immune-mediated OME. However, it seems possible that delayed type hypersensitivity may contribute to the pathogenic mechanism of OME.

Suppression of immune-mediated otitis media by T-suppressor cells.

Effects of mucosa-derived T-suppressor cells on induction of IgG-mediated otitis media with effusion (OME) were investigated in mice, since antigen-antibody reactions in the tympanic cavity are pathogenic mechanisms of OME. Splenic T-suppressor cells from strain C3H/HeN female mice, to which ovalbumin (OVA) had been orally administered, were transferred intravenously to syngeneic mice. The mice receiving the T-suppressor cells were immunized with OVA intraperitoneally and then injected with OVA into the tympanic cavity. Nine of ten control mice, to which splenic T cells from saline-fed mice were administered, developed OME, while OME was seen in only one of ten mice receiving splenic T cells from OVA-fed mice. The results show that IgG-mediated OME can be suppressed to a certain extent by the induction of antigen-specific, mucosa-derived, T-suppressor cells.