No abstract available.
Deoxyglucose* ; Shock*

To evaluate the role of capsular polysaccharide (CPS) as a virulence factor, the interaction of V. vulnificus with mouse peritoneal macrophages and serum, which are involved in the clearance of bacteria from blood and other tissues, were examined. In this study, MO6-24/0 (wild strain; hemolysin- and capsule-positive), MO6-24/I' (acapsular spontaneous mutant), CVD 752 (acapsular transposon mutant), and CVD 707 (hemolysin-negative and capsule-positive mutant) were used. The strain with CPS (MO6-24/0 and CVD 707) were more resistant to phagocytosis by mouse peritoneal macrophages compared with acapsular strains (MO6-24/T and CVD 752), and the resistance to phagocytosis was not changed by serum opsonin in the capsular strains. Acapsular strains were more susceptible to serum bactericidal activity than the capsular strains through the classical complement pathway. MO6-24/0 strain were detected in blood, spleen, liver and lung at 4 hours after intraperitoneally infection, whereas CVD 752 were not detected. All tested strains could induced the transcription of inflammatory cytokine gene such as IL-1, IL-6, IL-10 and TNF-u, and their inductions were not decreased by cytochalasin B treatment. This results demonstrate that CPS of V. vulnificus plays an important role in V. vulnificus infection through interfering nonspecific host defense system such as blood clearance and phagocytosis.
Animals ; Bacteria ; Complement Pathway, Classical ; Cytochalasin B ; Interleukin-1 ; Interleukin-10 ; Interleukin-6 ; Liver ; Lung ; Macrophages, Peritoneal ; Mice ; Phagocytosis ; Spleen ; Vibrio vulnificus* ; Vibrio* ; Virulence*

No abstract available.
Calcium* ; Cholera Toxin* ; Cholera* ; Cytoplasm* ; Humans* ; T-Lymphocytes*

This study was conducted in order to assess whether the form of the shade guide affects in deciding the color of the teeth using the shade guide. Eight shade light cured composite resins (Esthet-X, Dentsply, Milford, USA) were used in this study. Shade guides including the model of maxillary central incisors, teeth-form shade guide, doughnut form shade guide, and shade guide with perforated gray shield were prepared with eight shade composite resins and provided the codes randomly. After arranging the models of teeth, 19 dentists working at the clinic of the Dentistry of Chonnam University Hospital and 65 students of college of dentistry, Chonnnam University selected the shade guides corresponding to the color of each tooth on the gray board under the D65 standard illuminant. B1 shade showed highest accuracy of about 95% among all shade guides of 3 forms applied to the test and regardless of observer, tooth form shade guide showed the highest accuracy (p < 0.05), and the doughnut form showed the lowest accuracy (p < 0.05). At the time of deciding on the color of the teeth using the shade guides as a result of above, the forms of the shade guides can affect the accuracy, and it suggests that the development of the diversified forms of shade guides, which may obtain more accurate results, is required.
Composite Resins ; Dentistry ; Dentists ; Humans ; Incisor ; Jeollanam-do ; Tooth

The establishment of effective preventive measure against V. vulnificus septicemia is urgently required. It was reported that V. vulnificus osmotically shocked by distilled water lost viability rapidly but regain viability after appropriate resuscitation (RS) procedure. But V. vulnificus was reported to be completely killed when osmotically shocked in the presence of ethylenediaminetetraacetic acid (EDTA). This study was carried out to uncover the bactericidal mechanism of osmotic shock and the mechanism of potentiation of osmotic shock by EDTA. When about 2.0 x 10(7) CFU/ml of V. vulnificus were inoculated in distilled water, the number of viable cells abruptly decreased to 2.5 x 10(3) CFU/ml in 1 min. and slowly thereafter to 1.0 x 10(1) CFU/ml in 5 min. After RS, there was a increase in the number of surviving bacteria by 10(3) to 10(4) fold. When the bacteria were inoculated in 1 mM EDTA solution, osmotic concentration of which is about 30 mEq./1, no colony could be observed even in 1 minute. The turbidity decreased abruptly as soon as the bacteria were inoculated in distilled water or in the 1 mM EDTA solution, but rather slowly thereafter. When V. vulnificus whose cellular constituents were labeled with 3H-L-amino acid mixture was inoculated in distilled water or in the 1 mM EDTA solution, about 35% of the whole cell radioactivity was released in the 1 mM EDTA solution in 30 sec while about 6% of the whole cell radioactivity was released to the supernatant in distilled water in 5 minutes. The cell surface hydrophilicity decreased significantly by osmotic shock. The decrease was more significant when the bacteria were inoculated in 1 mM EDTA solution than in distilled water. Bacterial cell volume analysis with a flow cytometer revealed that the osmotic shock balloons V. vulnificus. The increase in the cell volume was more prominent in 1 mM EDTA solution. When the cytoplasmic RNA content in the osmotically shocked bacteria was measured by a flow cytometer, the frequency of the cells with decreased RNA content increased after osmotic shock, and the degree of increase was more prominent in 1 mM EDTA solution. Number of non-staining cells also increased after osmotic shock, and the degree of increase was more prominent in the 1 mM EDTA solution. To see whether the susceptibility to osmotic shock is unique to V. vulnificus, bactericidal kinetic curves of other Vibrio species were observed after inoculating in distilled water. V. cholerae and V. mimicus were more resistant to the osmotic shock than V. vulnificus. V. parahaemolyticus, V. furnissii, V. fluvialis, V. damsela, and V. harveyi showed similar susceptibility to osmotic shock as V. vulnificus. V. alginolyticus and V. hollisae were more susceptible than V. vulnificus. The concentration of NaCl in culture media influenced the susceptibility of V. vulnificus to osmotic shock. V. vulnificus grown in 0.5% NaCl was more resistant to the osmotic shock than that grown in 2.5% NaCl. Taken together, it was concluded that osmotic shock causes leakage of the cytoplasmic contents(ribosomes etc.). And EDTA was supposed to quantitatively potentiate the bactericidal effect of the osmotic shock. Susceptibility to osmotic shock was influenced by the osmolarity of culture media and appeared to be a phenotypic property of V. vulnificus.
Bacteria ; Cell Size ; Cholera ; Culture Media ; Cytoplasm ; Edetic Acid ; Hydrophobic and Hydrophilic Interactions ; Osmolar Concentration ; Osmotic Pressure* ; Radioactivity ; Resuscitation ; RNA ; Sepsis* ; Shock ; Vibrio vulnificus* ; Vibrio* ; Water

Toll-like receptors (TLRs) are pattern recognition receptors (PRRs) expressed in a wide spectrum of cell types that recognize distinctive ligands and subsequently activate adaptive immune responses. TLR ligands are considered a promising target for development of immunomodulatory agents. Extensive clinical investigations are currently underway to develop TLR ligands-based non-specific immunostimulants and vaccine adjuvants. It has been well accepted that cancer cells develop a strategy to avoid host immune responses by producing inhibitory molecules. In addition, tumor-associated antigens are often not strong enough to induce effective anti-cancer immune responses. In this context, immunostimulants or adjuvants are critically required for more effective cancer immunotherapies. Here, we discuss recent progresses in the field of cancer immunotherapy under special emphasis on the TLR ligands as a component of immunostimulatory agents.
Adjuvants, Immunologic ; Immunotherapy ; Ligands ; Receptors, Pattern Recognition ; Toll-Like Receptors

Mucosal vaccination, capable of inducing protective immune responses both in the mucosal and systemic immune compartments, has many advantages and is regarded as a blue ocean in the vaccine industry. Mucosal vaccines can offer lower costs, better accessability, needle-free delivery, and higher capacity of mass immunizations during pandemics. However, only very limited number of mucosal vaccines was approved for human use in the market yet. Generally, induction of immune responses following mucosal immunization requires the co-administration of appropriate adjuvants that can initiate and support the effective collaboration between innate and adaptive immunity. Classically, adjuvant researches were rather empirical than keenly scientific. However, during last several years, fundamental scientific achievements in innate immunity have been translated into the development of new mucosal adjuvants. This review focuses on recent developments in the concepts of adjuvants and innate immunity, mucosal immunity with special interest of vaccine development, and basic and applied researches in mucosal adjuvant.
Achievement ; Adaptive Immunity ; Cooperative Behavior ; Humans ; Immunity, Innate ; Immunity, Mucosal ; Immunization ; Mass Vaccination ; Pandemics ; Vaccination ; Vaccines

The motile marine bacterium, Vibrio vulnificus has a total of six flagellins. Flagellin is a structural component of flagellar filament in various locomotive bacteria and is the ligand of Toll-like receptor 5 (TLR5). TLRs, highly expressed on various types of cells including dendritic cells (DCs), recognize invading microorganisms and finally trigger host immune responses. In this study, we prepared all of six recombinant flagellin proteins and assessed the effect of six flagellins on IL-8 activation through TLR5 recognition. Although showed different activities, five out of the six flagellins stimulated significant IL-8 activation. We also investigated the immunomodulatory roles of Vv-FlaB, the crucial building block of V. vulnificus flagellar filament, on human dendritic cells. Treatment of immature DCs with Vv-FlaB resulted in an increased expression of co-stimulatory molecules and induced strong allo-T cell proliferative activities of the DCs. These results show that the Vv-FlaB may serve an epochal immune adjuvant possessing effective immunomodulatory activities.
Bacteria ; Dendritic Cells* ; Flagellin* ; Flow Cytometry ; Humans* ; Interleukin-8 ; Toll-Like Receptor 5 ; Vibrio vulnificus* ; Vibrio*

Selecting an appropriate antigen with optimal immunogenicity and physicochemical properties is a pivotal factor to develop a protein based subunit vaccine. Despite rapid progress in modern molecular cloning and recombinant protein technology, there remains a huge challenge for purifying and using protein antigens rich in hydrophobic domains, such as membrane associated proteins. To overcome current limitations using hydrophobic proteins as vaccine antigens, we adopted in silico analyses which included bioinformatic prediction and sequence-based protein 3D structure modeling, to develop a novel periodontitis subunit vaccine against the outer membrane protein FomA of Fusobacterium nucleatum. To generate an optimal antigen candidate, we predicted hydrophilicity and B cell epitope parameter by querying to web-based databases, and designed a truncated FomA (tFomA) candidate with better solubility and preserved B cell epitopes. The truncated recombinant protein was engineered to expose epitopes on the surface through simulating amino acid sequence-based 3D folding in aqueous environment. The recombinant tFomA was further expressed and purified, and its immunological properties were evaluated. In the mice intranasal vaccination study, tFomA significantly induced antigen-specific IgG and sIgA responses in both systemic and oral-mucosal compartments, respectively. Our results testify that intelligent in silico designing of antigens provide amenable vaccine epitopes from hard-to-manufacture hydrophobic domain rich microbial antigens.
Animals ; Cloning, Molecular ; Computational Biology ; Computer Simulation* ; Epitopes ; Epitopes, B-Lymphocyte ; Fusobacterium nucleatum* ; Fusobacterium* ; Hydrophobic and Hydrophilic Interactions ; Immunoglobulin A, Secretory ; Immunoglobulin G ; Membrane Proteins ; Mice ; Periodontitis ; Solubility ; Vaccination

Among the exotoxins produced by V. vulnificus, hemolysin (HS) has been reported to be the most potent one. To investigate the factors up- or down-regulating HS production in the context of pathogenesis, we observed the effects of salinity or/and temperature shifting, glucose, and acidic pH on the production of HS by V. vulnificus C7184 strain in vitro. Significantly more HS was produced when V. vulnificus was cultured in 0.9% salinity and 37 degrees C than in 2.5% and 25 degrees C. When the culture condition reflecting natural habitat of V. vulnificus (2.5% salinity and 25degrees C) was changed into that reflecting human body (0.9% salinity and 37 degrees C), 2.5 fold or more HS was produced than in the V. vulnificus being cultured continuously in 0.9% NaCl at 37 degrees C. This result suggests that V. vulnificus somehow recognizes the shifting in salinity and temperature and stimulate HS production. Glucose addition in the culture medium resulted in a dose- dependent decrease in the HS production. Glucose itself and acidic pH resulting from its metabolism both appeared to inhibit the HS production. Glucose in itself had more dominant role in suppressing the HS production than the lowered pH accompanying the metabolism of glucose. This result suggests that HS production is down-regulated in the presence of glucose and under environmental acidic pH.
Ecosystem ; Exotoxins ; Glucose* ; Human Body ; Hydrogen-Ion Concentration ; Metabolism ; Salinity* ; Vibrio vulnificus* ; Vibrio* ; Virulence