The whole cell extract of Helicobacter pylori strain 26695 was treated with the hemin-agarose resin and the bound fraction was analyzed by 2-Dimensional electrophoresis. The 2-D-PAGE-displayed spots were eluted and analyzed by matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). Among the 120 spots processed, 94 protein spots were identified to represent 58 genes. Forty-five protein spots that represented thirty-four genes were newly identified in this study, including iron-containing proteins and hemin-containg proteins such as fumarate reductase, iron-sufur subunit(FrdB), ribonucleoside diphosphate reductase, beta subunit (NrdB), glutamyl-tRNA reductase (HemA), nikel-cobalt-cadnium resistance protein (NccB), and porphobilinogen deaminase (HemC).
Chromatography, Affinity* ; Electrophoresis ; Helicobacter pylori* ; Helicobacter* ; Hydroxymethylbilane Synthase ; Mass Spectrometry ; Oxidoreductases ; Proteome* ; Ribonucleoside Diphosphate Reductase ; Succinate Dehydrogenase

The nucleotide sequence of pZMO1, a small cryptic plasmid of Zymomonas mobilis ATCC10988 was determined. Analysis of 1,680 bp of sequence revealed 69% identity with Shigella sonnei plasmid, pKYM and 61% identity with Nostoc sp. ss DNA replicating plasmid. Analysis of a deduced amino acid sequence of an orf of pZMO1 revealed 75% identity and 90% similarity with the repA gene of Synechocystis sp. plasmid pCA2.4. The upstream region of the repA gene of pZMO1 possesses six directed repeat sequences and two inverted repeat sequences at downstream of the IR consensus sequence of nick region of rolling circle replication (RCR) plasmid. A typical terminator hairpin structure was found at the downstream region of repA gene. Degradation of single-stranded plasmid DNA by S1 nuclease was detected by Southern hybridization. It suggests that pZMO1 replicates by a rolling circle mechanism in Z. mobilis ATCC10988 cells.
Amino Acid Sequence ; Animals ; Base Sequence ; Consensus Sequence ; DNA ; Ecthyma, Contagious ; Inverted Repeat Sequences ; Nostoc ; Plasmids* ; Shigella sonnei ; Synechocystis ; Zymomonas*

No abstract available.
Hypertension, Renovascular*

The component of Helicobacter pylori responsible for mouse death was identified and partially characterized. Mice that were injected with H. pylori cell lysate showed pathological changes such as decreased activity, diarrhea, mild convulsion, dramatic decline of body temperature, and even death. In order to identify the lethal factor, recently isolated H. pylori strain 335 and old culture (for 10 years) of H. pylori strain 51 were used. LD50 of the cell lysate of H. pylori 335 and 51 were 338 microgram and 985 microgram, respectively, meaning the long passage of H. pylori in the laboratory might have decreased the lethal activity in the lysate. Mouse lethal activity disappeared by either treatment of cell lysate with proteinase K or heating cell lysate at 60 degrees C for 30min. Mutation analysis and genetic complementation study revealed that active urease of H. pylori is the mouse lethal factor. The recombinant H. pylori urease expressed in Escherichia coli showed si milar lethal activity.
Animals ; Body Temperature ; Complement System Proteins ; Diarrhea ; Endopeptidase K ; Escherichia coli ; Heating ; Helicobacter pylori* ; Helicobacter* ; Hot Temperature ; Lethal Dose 50 ; Mice* ; Seizures ; Urease*

Helicobacter pylori (H. pylori) may be one of the most common pathogen in gastrointestinal tract. Although several recent articles have reported a decline in the prevalence of H. pylori infection in both children and adults over the last several years, H. pylori infection usually occurs early in life and persists for a long time. The role of H. pylori in some digestive diseases, such as gastritis, ulcer and gastric cancer has been well established. And the possible role of H. pylori as a trigger for some extraintestinal diseases in children and adults has been considered in the last year. H. pylori infection might be associated with refractory iron deficiency anemia, idiopathic thrombocytopenic purpura, growth retardation and obesity etc, directly or indirectly. Most of the studies are classified as epidemiological, clinical researches about effects on extraintestinal manifestations after eradication, or case reports. This review describes the possibility of association with several extraintestinal diseases and H. pylori infection and their possible mechanisms based on reported studies in the world and our several studies, even though there are still many conflicting results about that.
Adult ; Anemia, Iron-Deficiency ; Child ; Gastritis ; Gastrointestinal Tract ; Helicobacter ; Helicobacter pylori ; Humans ; Obesity ; Prevalence ; Purpura, Thrombocytopenic, Idiopathic ; Stomach Neoplasms ; Ulcer

As part of an initial inquiry into the function of the outer membrane proteins (OMPs) of Helicobacter pylori Korean strain 51, we have conducted an extensive proteome analysis via quadrupole time of flight (Q-TOF) mass spectrometry (MS). Fifty one OMPs of H. pylori were purified using sarcosine and resolved via two-dimensional electrophoresis with immobilized pH gradient strips. The most abundant proteins were observed in the alkaline pI regions (6.0~11.0) at molecular masses between 10~100 KDa. Here, 15 spots were identified, representing 9 types of genes (KHP0852, KHP0853, KHP1353, KHP1017, KHP0172, KHP0076, KHP0617, KHP1069, KHP0614) from the sarcosin-insoluble fraction of H. pylori 51. These may be employed in the characterization of the OMPs of H. pylori 51, which will help to identify new potential target proteins for vaccine development and drug therapy.
Electrophoresis ; Helicobacter ; Helicobacter pylori ; Mass Spectrometry ; Membrane Proteins ; Membranes ; Proteins ; Proteome ; Proton-Motive Force ; Sarcosine ; Sprains and Strains

Johne's disease (JD) is a chronic, debilitating disease of ruminants including cows, and is caused by Mycobacterium avium subsp. paratuberculosis (MAP). MAP is not only important in animal husbandry, but also in public health as it is associated with the onset of Crohn's disease, a chronic inflammatory bowel disease in humans. JD, like other mycobacterial diseases including tuberculosis, is classified into different stages based on the progression of infection. In addition, development of diagnostic assays that can distinguish between subclinical and clinical stages of JD is essential to control mycobacterial infection by providing an effective treatment. For the development of novel diagnostic methods of JD, it is important to investigate and understand the mRNA expression of the various immune markers in individuals at each stage of infection. In this study, we measured the levels of Th1-type chemokines, CXCR3, CCL4, CCL5, CXCL9, CXCL10, and CXCL11 in MAP-infected bovine blood by interferon (IFN)-γ release assay (IGRA) using IFN-γ as an alternative biomarker. The association of mRNA expression patterns of these chemokines with the MAP infection stages was analyzed and IFN-γ, CCL5, and CXCL10 were found to be significantly upregulated compared to IFN-γ, the biomarker used in IGRA. Our results further indicate that IFN-γ levels significantly increased in individuals with MAP-specific antibody, and CCL5 and CXCL10 levels significantly increased in those with MAP DNA. In particular, CCL5 was significantly upregulated in individuals, in which both MAP-specific antibody and MAP DNA were detected, but the expression of CXCL10 was specifically elevated in MAP DNA-detected individuals without MAP-specific antibody.
Animal Husbandry ; Animals ; Biomarkers ; Cattle* ; Chemokines* ; Crohn Disease ; DNA ; Gene Expression* ; Humans ; Inflammatory Bowel Diseases ; Interferons ; Mycobacterium avium subsp. paratuberculosis ; Mycobacterium avium* ; Mycobacterium* ; Paratuberculosis ; Public Health ; RNA, Messenger ; Ruminants ; Transcriptome* ; Tuberculosis

BACKGROUND: The problems of coronary stent thrombosis and restenosis still remain to be solved.The glycoprotein IIb/IIIa receptor blocker, Abciximab (ReoPro), plays important roles in the treatment of high-risk patient with acute platelet-rich thrombus and in the inhibition of smooth muscle cell proliferation. The aim of this study was to determine whether the use of ReoPro-coated stents could reduce the neointimal formation in a porcine coronary stent restenosis model. METHODS: ReoPro was coated on the surface of stent by means of plasma polymerization followed by chemical grafting. Stent overdilation injury was performed with control bare stent (Group I, n=13), and ReoPro-coated stents (Group II, n=14). Follow-up quantitative coronary angiogram was performed at 4 weeks after stenting and histopathologic assessment were compared in both groups. RESULTS: The diameter stenosis by QCA between two groups was significantly higher in Group I (23+/-5 % vs. 15+/-7 %, p=0.003). On histopathologic examination, no in-stent thrombus was observed. The percent area stenosis was significantly higher in Group I than in Group II (48+/-17 % vs. 30+/-16 %, p=0.01). The area of neoinima was larger in Group I than in Group II (3.2+/-1.2 mm2 vs. 2.0+/-1.0 mm2, p=0.01). By immunocytochemistry, proliferation cell nuclear antigen indices were higher in Group I (4.2+/-2.1 %, vs 2.4+/-1.8 % p=0.03). CONCLUSION: The ReoPro-coated stent is safe and effective in the prevention of in-stent thrombus and restenosis, which may be related with the inhibition of platelet thrombus and neointimal cell proliferation.
Blood Platelets* ; Cell Proliferation ; Constriction, Pathologic ; Follow-Up Studies ; Glycoproteins* ; Humans ; Immunohistochemistry ; Myocytes, Smooth Muscle ; Neointima ; Plasma ; Polymerization ; Polymers ; Stents* ; Thrombosis ; Transplants

Helicobacter pylori, a gram-negative bacterium, is a causative agent of gastroduodenal diseases of human. Human immune system produces harmful reactive oxygen species to kill this bacterium that locates the microaerophilic mucous layer. H. pylori harbors various antioxidant enzymes including SodB, KatA and AhpC to protect the oxygen toxicity. We removed the catalase gene (katA) from H. pylori 26695 genome, and the change of profile of the gene expression of the mutant was analyzed by high resolution 2-DE followed by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), tandem MS and microarray analysis. Eleven and 37 genes were upregulated and downregulated in the mutant respectively, either transcriptionally or translationally. Expression level of pfr and hp1588 that were decreased on protein level in the mutant was confirmed by RT-PCR analysis.
Catalase ; Gene Expression ; Genome ; Helicobacter pylori* ; Humans ; Immune System ; Mass Spectrometry ; Microarray Analysis ; Oxygen ; Proteome* ; Reactive Oxygen Species

It is commonly believed in the Western World that the more severe forms of gastroduodenal diseases like peptic ulcer are associated with infection by specific Helicobacter pylori strains classified as type I being considered to be more virulent than type II strains. However, in Korea, most of H. pylori isolates belong to type 1 strains regardless of virulence. Type I H. pylori strains differ from type II strains by the presence of the cag pathogenicity island (cag PAI) composed of a block of genes. In this study, the nucleotide sequence of cag PAI of the H. pylori Korean strain 51 was determined and compared with those of strains 26695 and J99 to assess the structural variation in the region and to evaluate its implication in the virulence of the H. pylori. The cag PAI of H. pylori strain 51 was smaller in size and in the number of constituting ORFs in comparison with 26695 and J99 strains. Although many cag orthologues were nearly identical one another with the similarity of 90% or more at the nucleotide and amino acid levels, there were some remarkable and significant differences in several cag genes among the three cag PAIs. Surprisingly, the percent similarities at amino acid level were lower than those at nucleotide level in one third of the ORFs. The two genes (cag7 and cagA) of strain 51 differed in sizes and deduced amino acid sequences from the corresponding genes of the other two strains. When comparing cagA ORF of H. pylori strain 51 with that of 8 non-Korean strains, phylogenetic tree revealed that the strain 51 formed a separate branch with the most far distances from the other strains except for a Japanese strain. The Cag7 protein of, strain 51 had a deletion in the repeat region II, suggesting a major change in the conformation and function of the protein.
Amino Acid Sequence ; Animals ; Asian Continental Ancestry Group ; Base Sequence ; Ecthyma, Contagious ; Genetic Variation ; Genomic Islands* ; Helicobacter pylori* ; Helicobacter* ; Humans ; Korea ; Open Reading Frames ; Peptic Ulcer ; Virulence* ; Western World