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Essential role of an adenylate cyclase in regulating <i>Vibrio vulnificus</i> virulence
Kim, Young Ran,Kim, Soo Young,Kim, Choon Mee,Lee, Shee Eun,Rhee, Joon Haeng Elsevier 2005 FEMS microbiology letters Vol.243 No.2
<P><B>Abstract</B></P><P><I>Vibrio vulnificus</I>, a halophilic estuarine bacterium, causes a fatal septicemia and necrotizing wound infection. To investigate the role of cAMP in <I>V. vulnificus</I> virulence regulation, an in-frame deletion mutant of the <I>cya</I> gene encoding adenylate cyclase was constructed. The <I>cya</I> null mutation resulted in a pleiotropic change of virulence phenotypes. The production of hemolysin and protease, the motility, and the cytotoxicity were decreased by the <I>cya</I> mutation. The defects in the <I>cya</I> mutant were functionally complemented <I>in trans</I> by a plasmid carrying the wild type <I>cya</I> allele. The <I>V. vulnificus cya</I> mutant exhibited a 100-fold increase in LD<SUB>50</SUB> to mice. The result indicates that cAMP plays an essential role in the global regulation of <I>V. vulnificus</I> virulence.</P>
Effect of Salinity, Temperature, and Glucose on the Production of Vibrio vulnificus Hemolysin
Kim, Hyun-Soo,Shin, Sung-Heui,Park, Hae-Ryoung,Lee, Shee-Eun,Kim, Choon-Mee,Kim, Soo-Young,Kim, Young-Ran,Lee, Hyun-Chul,Chung, Sun-Sik,Rhee, Joon-Haeng The Korean Society for Microbiology 2002 Journal of Bacteriology and Virology Vol.32 No.4
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 $37^{\circ}C$ than in 2.5% and $25^{\circ}C$. When the culture condition reflecting natural habitat of V. vulnificus (2.5% salinity and $25^{\circ}C$) was changed into that reflecting human body (0.9% salinity and $37^{\circ}C$), 2.5 fold or more HS was produced than in the V. vulnificus being cultured continuously in 0.9% NaCl at $37^{\circ}C$. 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.
Kim, Young-Ran,Kim, Choon-Mee,Rhee, Joon-Haeng The Korean Society for Microbiology 2003 Journal of Bacteriology and Virology Vol.33 No.1
Live Vibrio vulnificus is highly cytotoxic to host cells in vivo and in vitro. The two most representative cytotoxins, cytolytic hemolysin and elastolytic protease, have been regarded to play major roles in the cytotoxicity of V. vulnificus. To further determine roles of the two cytotoxins in V. vulnificus pathogenesis, we constructed a double mutant of vvhA and vvpE genes, encoding a hemolysin and a protease, respectively. However, the cytotoxicity and the $LD_{50}$ of a vvhA/vvpE double mutant showed no significant difference from those of the isogenic wild type strain. From these results, we came to speculate that yet unidentified, key cytotoxic factors should playa major role in the cytotoxic activity of V. vulnificus. The HeLa cells encountered with V. vulnificus became rounded, following detachment from the bottom of culture plate, and were killed eventually. However, the bacterial culture supernatant did not show any effect on the morphology and viability of HeLa cells. Also, no cytotoxicity could be noted when V. vulnificus was not allowed to contact with HeLa cells in the $Transwell^{(R)}$ system. Chloramphenicol, at lower concentration than minimum inhibitory concentration (MIC), decreased the cytotoxicity of a vvhA/vvpE double mutant to HeLa cells in a dose dependent manner. These results suggest that close encounter of V. vulnificus with host cells is a prerequisite to the cytotoxicity and that a yet unidentified virulence factor (s) should play an important role in the contact-dependent cytotoxicity.
Kim, Young Ran,Lee, Shee Eun,Kang, In-Chol,Nam, Kwang Il,Choy, Hyon E.,Rhee, Joon Haeng Oxford University Press 2013 The Journal of Infectious Diseases Vol.207 No.9
<P><I>Vibrio vulnificus</I>, a halophilic estuarine bacterium causing fatal septicemia and necrotic wound infection, is highly cytotoxic to eukaryotic cells. We have reported that RtxA1 toxin kills host cells only after they come into contact with bacteria and plays an essential role in the pathogenesis of <I>V. vulnificus</I>. This study was performed to elucidate the mechanism by which the RtxA1 toxin mediates the death of HeLa cells. By using confocal microscopy and immunoblot analysis, we show that the 501-kDa RtxA1 toxin is processed into 2 fragments after its secretion into host cells. The largerN-terminal fragment (RtxA1-N; approximately 370 kDa) remained at the host cell membrane, whereas the smaller C-terminal fragment (RtxA1-C; approximately 130 kDa) was internalized into the host cell cytoplasm. RtxA1-N is believed to polymerize and form pores at the host cell membrane and to induce an increase in necrotic volume related to calcium. The RtxA1 toxin caused an increase in the intracellular Ca<SUP>2+</SUP> concentration and the subsequent activation of JNK. The cell death mechanism occurred via calcium-dependent mitochondrial pathways, which caused calcium sequestration in the mitochondria, accompanied by irreversible mitochondrial membrane dysfunction and adenosine triphosphate depletion, and was later accompanied by the disruption of the integrity of the plasma membrane.</P>
Kim, Il-Ok,Kim, Mi-Ran,Shin, Jang-Hoon,Kim, Sun-Haeng korean Academy of Physical Therapy Rehabilitation 2020 Physical therapy rehabilitation science Vol.9 No.4
Objective: This study was conducted to provide basic data on physical function improvement activities of elderly care facilities by identifying the degree of physical improvement activities of elderly caregivers. Design: Descriptive correlational study. Methods: The instrument of this study consisted of 12 questions on the general characteristics of the subject. For the assessment of improvement in physical function activities, the measuring tool used for the elderly consisted of a total of 20 questions. For data analysis, Pearson's correlation coefficient & Spearman's rho and multiple regression were used. Results: The higher the age of the subjects, the lower their educational background (r=-0.273, p<0.05), and the higher the probability of having a religion (r=-0.258, p<0.05), the more stable the employment type (r=-0.333, p<0.05). The higher the level of education, the higher the monthly income (r=0.187, p<0.01), and the shorter the career (r=-0.204, p<0.05). The more stable employment, the more unstable duty (r=-0.245, p<0.05), and the more unstable work, the higher the monthly income (r=-0.206, p<0.05) and the longer the career, the higher the monthly income (r=0.247, p<0.05). The more stable the employment, the more activities to improve physical function were found (r=0.341, p<0.05), and the more unstable the duty, the more activities to improve physical function were found (r=0.321, p<0.05), and the higher monthly income, the less physical function improvement activities (r=-0.196, p<0.05). Conclusions: It is necessary to provide regular services by a dedicated physical therapist for physical function improvement activities in order to improve the quality of life of the elderly in the future.
Kim, Haeng Ran,Kim, Kyung Mi,Lee, Ju-Hye,Yoo, Sang-Ho,Lee, Soh Min,Kim, Kwang-Ok Korean Society of Food Science and Technology 2016 Food Science and Biotechnology Vol.25 No.5
Long steeping process of waxy rice involved in Gangjung production is essential for obtaining good quality Gangjung, but it causes many related problems. The objective was to examine physiochemical properties of waxy rice flour and expansion properties of Gangjung manufactured using waxy rice flour treated with protease (P-flour), ${\alpha}$-amylase (A-flour), or in combination of the two (PA-flour) over different durations. While crude protein and reducing sugar contents of P-flour and A-flour differed depending on enzyme treatment time, it was not observed for PA-flour. Gangjung made with P-flour or A-flour could show similar expansion rate to that of Gangjung made with waxy rice flour steeped under the optimal condition, however, Gangjung made with PA-flour showed lower expansion rate than of the optimal one. These findings indicate that treatment of waxy rice flour with either protease or ${\alpha}$-amylase, but not both, could reduce the steeping time involved in Gangjung production.
Relationship of Specific Microbial Growth and TBARS Value in Radiation-Sterilized Raw Ground Pork
Kim, Jae-Kyung,Jo, Cheo-Run,Kim, Hyun-Joo,Lee, Kyong-Haeng,Kim, Yeung-Ji,Byun, Myung-Woo The Korean Society of Food Science and Nutrition 2004 Preventive Nutrition and Food Science Vol.9 No.4
Sterilized raw ground pork was inoculated with Pseudomonas aeruginosa (PA) and Lactobacillus casei (LC) to investigate the relationship between microbial growth and 2-thiobarbituric acid reactive substances (TBARS) values. The analyses including microbial growth, pH, and TBARS values were performed during 3 weeks of storage at room temperature $(20^{\circ}C)$. The radiation-sterilized control sample did not show any microbial growth, but the samples inoculated at different levels (diluted twice vs non-diluted) exhibited differences until 1 week. However, the difference disappeared at weeks 2 and 3. The pH of raw ground pork inoculated with PA increased, but that of LC decreased. The pH of non-inoculated samples increased slightly after storage. The TBARS values in non-inoculated and LC inoculated with pork increased, but TBARS remained unchanged in samples inoculated with PA after 1 week. Results indicated that the microbial growth level and strains can influence the TBARS value of raw ground pork. Thus, it is important to use samples exposed to the same microbial conditions to compare the oxidation of lipids in meat samples.