Roles of major oligosaccharides on Cry j 1 in human immunoglobulin E and T cell responses.

BACKGROUND: We have demonstrated that carbohydrates in Cry j 1, the major allergen of Cryptomeria japonica pollen, play a major role in promoting Cry j 1-specific Th2 response. However, little is known as to whether the carbohydrates directly participate in allergic responses. OBJECTIVE: We sought to determine whether Cry j 1-related oligosaccharides function as IgE and/or T cell epitopes. In addition, the regulatory effect of Cry j 1-related oligosaccharide on Cry j 1-specific T cell responses was investigated. METHODS: Two monovalent oligosaccharides largely found on Cry j 1, Manalpha1-6(Manalpha1-3)(Xylbeta1-2)Manbeta1-4GlcNAcbeta1-4(Fucalpha1-3)GlcNAc (M3FX), and GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1-3)(Xylbeta1-2)Manbeta1-4GlcNAcbeta1-4(Fucalpha1-3)GlcNAc (GN2M3FX) were prepared. Manalpha1-2Manalpha1-6(Manalpha1-2Manalpha1-3)Manalpha1-6(Manalpha1-2Manalpha1-2Manalpha1-3)Manbeta1-4GlcNAcbeta1-4GlcNAc (M9A) was used as control. Competitive inhibition ELISA for Cry j 1-specific IgE was performed using these oligosaccharides as inhibitors. In addition, T cell lines specific for Cry j 1 or purified protein derivative of Mycobacterium tubecurosis (PPD) were established, and cellular responses against these oligosaccharides were investigated in the presence or absence of the respective antigens. RESULTS: Overall, neither M3FX nor GN2M3FX displayed inhibitory effect on the binding between IgE and Cry j 1. In addition, M3FX did not by itself stimulate Cry j 1 or PPD-specific T cells. However, M3FX significantly inhibited Cry j 1-induced proliferation and IL-4 production in Cry j 1-specific T cells. Such an inhibitory effect was not seen in PPD-specific T cell responses. CONCLUSION: These results suggest that Cry j 1-related oligosaccharides are not major epitopes for IgE or T cells. However, these oligosaccharides have a novel potential to inhibit Cry j 1-specific T cell responses selectively.

Dantaxusins A and B, two new taxoids from Taxus yunnanensis.

Two new taxane diterpenes, dantaxusin A [5 alpha-cinnamoyloxy-2 alpha,7 beta,13 alpha-triacetoxy-2(3-->20)abeo-taxa-4(20),11-diene-9,10-dione (1)] and dantaxusin B [5 alpha-cinnamoyloxy-9 alpha-hydroxy-10 beta,13 alpha-diacetoxytaxa-4(20),11-diene (2)], were isolated from an ethanol extract of the aerial parts of Taxus yunnanensis along with taxuspine B, 2-deacetoxytaxinine J, taxuyunnanine C, taxinine B, taxuspine C, and taxinine NN-4. The structures of 1 and 2 were established on the basis of 1D and 2D NMR and HRMS spectroscopic methods.

Roles of carbohydrates on Cry j 1, the major allergen of Japanese cedar pollen, in specific T-cell responses.

BACKGROUND: Carbohydrates expressed on allergens are known to be important for allergenicity. However, little is known about whether the carbohydrates drive the T(H)2 response. OBJECTIVE: We sought to determine a role for carbohydrates expressed on Cry j 1, which is the major allergen of Cryptomeria japonica pollen and causes the most prevalent pollinosis in Japan, in in vitro cellular responses. METHODS: Carbohydrates on Cry j 1 were destroyed by periodate-oxidation under mild conditions. Proliferative responses and cytokine productions against native, periodate-treated, and mock-treated Cry j 1 were compared in peripheral blood mononuclear cells, Cry j 1-specific T-cell lines, and clones from patients with Japanese cedar pollinosis. RESULTS: We found that peripheral blood mononuclear cells from patients with Japanese cedar pollinosis displayed a significant decrease in proliferation and IL-5 production in response to periodate-treated Cry j 1 in comparison with native and mock-treated Cry j 1. Decreased proliferative responses against periodate-treated Cry j 1 were also seen in polyclonal T-cell lines, and the responses showed a heterogeneity. In addition, Cry j 1-specific CD4+ T-cell clones also displayed a significant decrease in proliferation and IL-4 and IL-5 production-but not IFN-gamma production-in comparison with the control antigens. However, most of the clones showed decreased but positive proliferative responses against periodate-treated Cry j 1. Blockade of the mannose receptor had no effect on cellular responses. CONCLUSION: The results suggest that carbohydrates on Cry j 1 play a major role in promoting Cry j 1-specific T(H)2 response in vitro, though they are not major targets as T-cell epitopes.

Cytotoxity of non-alkaloidal taxane diterpenes from Taxus chinensis against a paclitaxel-resistant cell line.

Seven taxane diterpenes were isolated from the EtOH extract of the aerial parts of Taxus chinensis, and evaluated for cytotoxicity against nine human cell lines, including a beta-tublin mutant resistant to paclitaxel. Compound 2, a non-alkaloid-type taxane diterpene, showed significant cytotoxicity in most cell lines, and notably, equipotent against both parental and beta-tublin mutant tumor cell lines.

Topical gentian violet for cutaneous infection and nasal carriage with MRSA.

This study describes a potential effect of topical gentian violet on cutaneous infection and nasal carriage with methicillin-resistant Staphylococcus aureus (MRSA). 0.5% gentian violet was used in 28 cases of skin lesions once a day, while a 0.3% solution was applied on the nasal vestibules of nine cases twice a day. The period for eradication in the 28 skin cases was 9.1 +/- 6.0 days. It was 15.3 +/- 9.0 days for the nine nasal lesions. The minimal inhibitory concentration (MIC) of gentian violet against MRSA from the four isolated strains was 0.0225 +/- 0.0096 microg/mL. No adverse reactions occurred throughout the study. It is suggested that gentian violet may be potentially effective against MRSA.

Purification, identification, and cDNA cloning of Cha o 2, the second major allergen of Japanese cypress pollen.

The second major allergen of Chamaecyparis obtusa (Japanese cypress) pollen, Cha o 2, has been purified and its cDNA cloned. Of patients with pollinosis caused by C. obtusa, 82.5% produce IgE antibodies which react with purified Cha o 2. The purified protein has a molecular mass of 46 kDa and its 12 N-terminal amino acid sequence displays a high homology with that of Cry j 2, the second major allergen of Cryptomeria japonica pollen. cDNA clones coding for Cha o 2 have been isolated using Cry j 2 cDNA as a probe. Cha o 2 cDNA clones were sequenced and found to code a putative 50-residue signal sequence and a 464-residue mature protein with a molecular weight of 50 kDa. Two possible N-linked glycosylation sites were found in the sequence. The deduced amino acid sequence of Cha o 2 shows 74.3% identity with that of Cry j 2. In its primary structure, Cha o 2 shows significant identity with those of the polygalacturonases of avocado, tomato, and maize as well as Cry j 2.

Bruceanols G and H cytotoxic quassinoids from Brucea antidysenterica.

Two new quassinoids, bruceanols G [1] and H [4], were isolated from Brucea antidysenterica, and their structures were elucidated by spectral evidence and chemical transformation. Bruceanol G exhibited significant cytotoxicity against the COLO-205 and KB neoplastic cell lines with ED50 values of 0.44 and 0.55 microM, respectively.

Inhibitory effects of quassinoids on Epstein-Barr virus activation.

Short-term in vitro assays for tumor promoters and anti-tumor promoters (Epstein-Barr virus activation test) were carried out for 45 quassinoids. As a result, some quassinoids showed potent activity, more than 50% inhibition at a molar ratio of 1:1 (TPA/quassinoids). These results led to the following structure-activity relationships: (1) a methyleneoxy bridge and side chain enhance the activity and (2) a sugar moiety reduces the activity.

Bruceosides D, E, and F, three new cytotoxic quassinoid glucosides from Brucea javanica.

Three new quassinoid glucosides, bruceosides D [1], E [2], and F [3], were isolated from Brucea javanica, and their structures were elucidated by spectral evidence and chemical transformation to known compounds. Compounds 1-3 show selective cytotoxicity in the leukemia and non-small cell lung, colon, CNS, melanoma, and ovarian cancer cell lines with log GI50 values in the range of -4.14 to -5.72.

[Clinical characterization of allergic patients sensitized to Chamaecyparis obtusa--using AlaSTAT system].

Clinical characterization of patients with allergy to Chamaecyparis obtusa (Cypress) pollen and their specific IgE antibody was investigated by using AlaSTAT assay, a new method for measurement of antigen-specific IgE antibody. Positive rate of antigen-specific antibody in patients with allergic pollinosis was 83.5% for Japanese cedar pollen and 80.0% for Chamaecyparis obtusa pollen. 76.4% of patients were sensitized both to Japanese cedar and Chamaecyparis obtusa pollen, however, 7.9% were sensitized only to the latter. As for clinical symptoms, duration of attack was short and severity was mild or moderate in the patients sensitized only to Chamaecyparis obtusa pollen. Patients who visited hospital in April, in that Chamaecyparis obtusa pollen disseminated in the air predominantly, showed significant elevation in AlaSTAT scores compared with patients who visited in February and March (p < 0.05).

Bruceanols D, E, and F three new cytotoxic quassinoids from Brucea antidysenterica.

Three new quassinoids, bruceanols D [1], E [2], and F [3], were isolated from Brucea antidysenterica, and their structures were elucidated by spectral evidence and chemical transformation. All of these compounds exhibited cytotoxicity against five human tumor cell lines, malignant melanoma (RPMI-7951), lung carcinoma (A-549), ileocecal adenocarcinoma (HCT-8), epidermoid carcinoma of the nasopharynx (KB), and medulloblastoma (TE-671), and against murine lymphocytic leukemia (P-388).

Dietary fish oil alters rat milk composition and liver and brain fatty acid composition of fetal and neonatal rats.

This study was performed to determine the effects of dietary fish oil rich in docosahexaenoic acid [22:6(n-3)] on the milk composition of maternal rats and tissue fatty acid composition of fetal and neonatal rats. The fatty acid composition of dam's milk was affected by the dietary fat. The level of 22: 6(n-3) in the milk of dams fed the fish oil-containing diet was severalfold higher than that of dams fed the diet that did not contain fish oil, and the levels of 22: 6(n-3) in the phospholipids of the livers and brains of fetuses and pups of the fish oil-fed group were higher than those in fetuses and pups of the group that did not receive fish oil. These findings suggest that a high level of 22:6(n-3) in the livers and brains of fetal and neonatal rats can be achieved by intake of fish oil by the dams.

Seasonal change in maximal expiratory flow-volume pattern in patients with Japanese cedar pollenosis.

Fifteen patients with Japanese cedar pollenosis were examined for lower airway function. Flow-volume patterns obtained from flow-volume and volume-time curves during the pollen season (March) and outside of the pollen season (June) were evaluated. In a previous report we classified maximal expiratory flow-volume (MEFV) curves in five patterns from A to E. In the present study, the patterns did not vary between the two periods except in one patient. Eleven patients out of 15 showed type E patterns, in which the flow-volume curve was concave along its entire course. In most of the patients with severe or moderate symptoms of allergic rhinitis only during the pollen season, the curve shifted to the right, but the parameters of the curves did not increase significantly outside of season. These findings suggest that patients with Japanese cedar pollenosis suffer from continuous latent peripheral airway obstruction. Extremely slight changes in the flow rate were detected by comparing the curves obtained during the two periods.

Molecular diversity of neuronal-type calcium channels identified in small cell lung carcinoma.

Using the polymerase chain reaction (PCR), we identified RNA transcripts for two distinct classes of neuronal-type voltage-gated Ca2+ channels (VGCC) in a prototypic small cell lung carcinoma (SCLC) cell line, SCC-9. Oligonucleotide primers were designed to encode amino acid sequences common to alpha 1-subunits of known neuronal VGCC classes. Sequencing of complementary DNA (cDNA) clones derived from two independent PCR products revealed that one corresponded to a brain class A VGCC fragment predicted to encode a P-type VGCC (insensitive to dihydropyridines and omega-conotoxin) characteristic of cerebellar Purkinje cells but not previously identified in humans. The second PCR product was identical (except for one conservative nucleotide difference) to a fragment of the class D VGCC of neurons and neuroendocrine cells, which encodes an L-type VGCC (sensitive to dihydropyridines). By Northern blot analyses, both cDNAs hybridized to messenger RNAs (mRNAs) obtained from SCC-9; class D hybridized additionally to human cerebral cortical mRNA, but neither hybridized to mRNA from the skeletal muscle cell line TE671. Although no cDNA corresponding to class B VGCC (N-type) was identified, SCLCs are known to express VGCC that are sensitive to omega-conotoxin and coprecipitate with 125I-labeled-omega-conotoxin when complexed with serum IgG from patients with the Lambert-Eaton myasthenic syndrome. The multiple classes of neuronal-type VGCC expressed in SCLC could conceivably have both unique and related antigenic determinants that may give rise to antineuronal autoimmune responses. This would account for a spectrum of paraneoplastic neurologic disorders including the Lambert-Eaton syndrome and subacute cerebellar degeneration.

Antitumor agents, 127. Bruceoside C, a new cytotoxic quassinoid glucoside, and related compounds from Brucea javanica.

Bruceoside C [5], a new quassinoid glucoside, and related compounds were isolated from Brucea javanica, and their structures were elucidated by spectral data. Bruceoside C showed potent cytotoxicities against KB, A-549, RPMI, and TE-671 tumor cells.

Antitumor agents, 90. Bruceantinoside C, a new cytotoxic quassinoid glycoside from Brucea antidysenterica.

Three cytotoxic, quassinoid glycosides, the new bruceantinoside C [1] and the known yadanziosides G [2] and N [3] were isolated from the stem of Brucea antidysenterica. The structures of 1-3 were determined from their spectral data.

[A case of vesicosigmoidal fistula secondary to diverticulitis of the colon].

A case of vesicosigmoidal fistula secondary to the diverticulitis of colon is reported. A 63-year-old man was admitted to our clinic with the chief complaint of pneumaturia. Cystoscopy revealed an edematous, papillomatous lesion at the left posterior bladder wall. Although multiple diverticulosis of descending and sigmoid colon were demonstrated by barium enema, the presence of a fistula from the intestine to bladder was not confirmed. The presence of fistula was confirmed by detecting the orally administered charcoal in the urine. Resection of sigmoid colon with partial cystectomy was performed. Removed specimen revealed multiple sigmoidal diverticula and a fistula from a sigmoidal diverticulum to bladder through a firm mass. Histological examination of mass demonstrated inflammatory changes. This was the 58th case of vesicosigmoidal fistula due to diverticulitis of colon reported in the Japanese literature.