BACKGROUND: Allergic inflammation was induced by activated Th2 lymphocytes, leading to IgE production and eosinophil activation. A Th2 disproportion was shown in atopic children soon after birth. During specific allergen stimulation, an increase of Th2 cells was observed in most cases. In this study, we prepared new screening "whole blood" system for searching the anti-atopic materials. Cytokine production and IgE secretion from whole blood system were assessed and we confirmed the results by using animal system. METHODS: Pathological features in NC/Nga mice are similar to those observed in human atopic dermatitis. Whole blood from NC/Nga mouse was stimulated by using TNCB (Th2 activator) or candidate materials of anti-atopic dermatitis, and the production of cytokines (IL-4, IL-12, and IFN-gamma) were measured by ELISA. In order to confirm the results of whole blood system, in vivo test was done by using NC/Nga mice. RESULTS: In whole blood system, LPS and extracts of green tea, hardy orange and onion induced the production of IL-12 and IFN-gamma while they reduced the production of IL-4. Also, LPS and extracts of onion reduced IgE production. Though atopic dermatitis was observed from a mouse stimulated with TNCB, it was not when a mouse was co-stimulated in LPS or extracts of onion. The results are same as those observed in whole blood system. CONCLUSION: Whole blood system was simple and speedy methods for searching a materials compared with the conventional high-cost animal system. And the results using whole blood system was proved to be reliable in our experiments for screening anti-atopic material. We expect that the system can be applied to other experiments for searching similar materials.
Animals ; Asthma ; Child ; Citrus sinensis ; Cytokines ; Dermatitis ; Dermatitis, Atopic ; Enzyme-Linked Immunosorbent Assay ; Eosinophils ; Humans ; Immunoglobulin E ; Inflammation ; Interleukin-12 ; Interleukin-4 ; Lymphocytes ; Mass Screening ; Mice ; Onions ; Parturition ; Tea ; Th2 Cells

No abstract available.
Contrast Media* ; Endothelium*

PURPOSE:Carotid endarterectomy may benefit patients with bilateral carotid stenosis by improving cerebrovascular hemodynamics of ipsilateral as well as contralateral cerebral hemispheres. We investigated cerebrovascular hemodynamics after carotid endarterectomy in patients with contralateral carotid occlusion by acetazolamide stress brain SPECT. MATERIALS AND METHODS: Subjects were 14 symptomatic patients (all men, mean age 66 yrs) with carotid stenosis (> 50%) with contralateral carotid occlusion. Acetazolamide stress Tc-99m ECD brain SPECTs were performed within 2 weeks before and after carotid endarterectomy using one day protocol. Cerebral blood flow (CBF) and cerebrovascular reserve (CVR) were assessed visually. In 12 patients, correlation between the patency of proximal anterior cerebral or anterior communicating arteries (A1/A-comm) and the improvement of CBF or CVR after endarterectomy was assessed. RESULTS: Preoperative SPECT showed reduced CBF in 2 ipsilateral and 10 contralateral hemispheres. CVR was reduced in 4 ipsilateral and 9 contralateral hemispheres. Of 12 hemispheres with reduced CBF, 2 hemispheres (16.7%) showed improvement of CBF after endarterectomy. However, reduced CVR was improved in all 4 ipsilateral and 7 of 9 (78%) of contralateral hemispheres after endarterectomy. Three of 4 with stenotic A1/A-comm and 4 of 8 with patent A1/A-comm had reduced contralateral CVR. Reduced contralateral CVR improved in all 3 patients with stenotic A1/A-comm and 3 of 4 with patent A1/A-comm. CONCLUSION: Acetazolamide stress brain SPECT demonstrated improvement of compromised cerebrovascular reserve in not only ipsilateral but also contralateral hemispheres of patients with contralateral carotid occlusion after carotid endarterectomy, and may, therefore, be useful for evaluating cerebral blood flow and cerebrovascular reserve after carotid endarterectomy.
Acetazolamide* ; Arteries ; Brain Ischemia ; Brain* ; Carotid Stenosis* ; Cerebrum ; Endarterectomy ; Endarterectomy, Carotid* ; Hemodynamics ; Humans ; Male ; Tomography, Emission-Computed, Single-Photon*

The cytolysin A (ClyA) is a 34 kDa pore-forming cytotoxic protein and expressed by some enteric bacteria including Salmonella typhi. This toxin is transported on the bacterial surface and secreted without posttranslational modification. Using the surface display of ClyA, the expression vectors for 193-aa immunogenic antigen of spike protein (termed S1E) from severe acute respiratory syndrome coronavirus (SARS-CoV) were constructed. The vectors carried a gene encoding S. typhi ClyA conjugated to S1E at the C terminus (termed ClyA-S1E) and asd gene in pGEM-T and pBR322, named pGApLCS1E and pBApLCS1E, respectively. An asd-mutated E. coli transformed with these vectors could grow without diaminopimelic acid (DAP), indicating that they were stably maintained in such mutants. ClyA-S1E recombinant proteins from these vectors were expressed on the surface of the attenuated S. typhimurium deficient of global virulence gene regulator, ppGpp. However, they did not show the hemolytic activity on the blood agar plate and cytotoxicity against HeLa cells. To examine whether bacteria expressing ClyA-S1E induced the immune response against S1E, S. typhimurium deficient of ppGpp and Asd was transformed with these vectors and orally immunized in mice. In the western blotting against GST-conjugated S1E using the immunized mouse sera, it was shown that the significant band was detected in the mouse serum by the bacteria transformed with pGApLCS1E but not with pBApLCS1E. It indicates that the immune response producing antibody was dependent on the expression level of ClyA-S1E. Therefore, ClyA delivery system can be used for SARS vaccine development.
Agar ; Animals ; Bacteria ; Blotting, Western ; Coronavirus ; Diaminopimelic Acid ; Enterobacteriaceae ; Genes, vif ; HeLa Cells ; Humans ; Mice ; Perforin ; Protein Processing, Post-Translational ; Recombinant Proteins ; Salmonella ; Salmonella typhi ; Severe Acute Respiratory Syndrome

The cag7 gene of Korean H. pylori strains was analyzed by RFLP to develop a discriminatory tool for genotyping clinical isolates. For this study, a total of 82 H. pylori strains were isolated from the patients; 27 strains from the patients with chronic gastritis, 26 from duodenal ulcer, and 29 from gastric cancer. Genomic DNA was isolated and subjected to PCR targeting entire ORF or the repeat regions I and II of cag7 gene. PCR products from entire ORF or repeat region I of cag7 gene were divided into two types. However, there was no difference in the length of PCR products from the repeat region II. By the PCR genotyping of the entire cag7 gene, genotypes A and B were established, which showed approximately 5,100 and 5,500 bp PCR products, respectively. The repeat region I showed approximately 600 or 1,000 bp DNA fragments by PCR. The length of cag7 gene was determined by the size variation in the repeat region I. In addition, RFLP analysis of the PCR products of cag7 gene showed 11 subtypes, based on the major bands. These findings illustrate that the genetic diversity of the repeat region I would serve a reliable target for the genotyping of the cag7 gene.
Animals ; DNA ; Duodenal Ulcer ; Ecthyma, Contagious ; Gastritis ; Genetic Variation ; Genotype ; Helicobacter pylori* ; Helicobacter* ; Humans ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length* ; Stomach Neoplasms

Low-abundance cellular proteins normally invisible on the standard two-dimensional SDS-polyacrylamide gel electrophoresis (2-DE SDS-PAGE) map must be enriched appropriately in order to be visualized and identified in cells or tissues. We applied proteins of H. pylori strain 26695 to a immobilized heparin-affinity resin, which has an affinity for nucleic acid-binding proteins, protein biosynthesis factors, and growth factors. The whole cell extract of H. pylori strain 26695 was fractionated by the heparin-agarose chromatography, and was analyzed by 2-DE. The 2-DE SDS-PAGE displayed spots after silver staining, which were identified by matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). Among the ca. 150 spots that were processed, 79 proteins representing 57 genes were identified. Eleven proteins were determined to be nucleic acid-associated. Eighteen proteins were newly identified in this study, including DNA topoisomerase I. These results may provide guidance for enriching low abundance proteins of H. pylori and contribute to the construction of a master protein map of H. pylori.
Chromatography* ; DNA Topoisomerases, Type I ; Electrophoresis ; Electrophoresis, Polyacrylamide Gel ; Helicobacter pylori* ; Helicobacter* ; Heparin* ; Intercellular Signaling Peptides and Proteins ; Mass Spectrometry ; Protein Biosynthesis ; Proteome ; Silver Staining