To understand the molecular mechanism involved in the survivability of cold-tolerant lactic acid bacteria was of great significance in food processing, since these bacteria play a key role in a variety of low-temperature fermented foods. In this study, the cold-stress response of probiotic Lactobacillus plantarum K25 isolated from Tibetan kefir grains was analyzed by iTRAQ proteomic method. By comparing differentially expressed (DE) protein profiles of the strain incubated at 10℃ and 37℃, 506 DE proteins were identified. The DE proteins involved in carbohydrate, amino acid and fatty acid biosynthesis and metabolism were significantly down-regulated, leading to a specific energy conservation survival mode. The DE proteins related to DNA repair, transcription and translation were up-regulated, implicating change of gene expression and more protein biosynthesis needed in response to cold stress. In addition, two-component system, quorum sensing and ABC (ATP-binding cassette) transporters also participated in cell cold-adaptation process. These findings provide novel insight into the cold-resistance mechanism in L. plantarum with potential application in low temperature fermented or preserved foods.
Li, Qianqian;Ni, Hong;Meng, Shan;He, Yan;Yu, Ziniu;Li, Lin
Journal of Microbiology and Biotechnology
/
v.21
no.12
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pp.1330-1335
/
2011
N-Acyl homoserine lactones (AHLs) serve as the vital quorum-sensing signals that regulate the virulence of the pathogenic bacterium Erwinia carotovora. In the present study, an approach to efficiently restrain the pathogenicity of E. carotovora-induced soft rot disease is described. Bacillus thuringiensis-derived N-acyl homoserine lactonase (AiiA) was projected onto the surface of Pseudomonas putida cells, and inoculation with both strains was challenged. The previously identified N-terminal moiety of the ice nucleation protein, InaQ-N, was applied as the anchoring motif. A surface display cassette with inaQ-N/aiiA was constructed and expressed under the control of a constitutive promoter in P. putida AB92019. Surface localization of the fusion protein was confirmed by Western blot analysis, flow cytometry, and immunofluorescence microscopy. The antagonistic activity of P. putida MB116 expressing InaQ-N/AiiA toward E. carotovora ATCC25270 was evaluated by challenge inoculation in potato slices at different ratios. The results revealed a remarkable suppressing effect on E. carotovora infection. The active component was further analyzed using different cell fractions, and the cell surface-projected fusion protein was found to correspond to the suppressing effect.
Acute respiratory virus infectious diseases are a growing health problem, particularly among children and the elderly. Much effort has been made to develop probiotics that prevent influenza virus infections by enhancing innate immunity in the respiratory tract until vaccines are available. Lactobacillus plantarum GB-LP2, isolated from a traditional Korean fermented vegetable, has exhibited preventive effects on influenza virus infection in mice. To identify the molecular basis of this strain, we conducted a whole-genome assembly study. The single circular DNA chromosome of 3,284,304 bp was completely assembled and 3,250 protein-encoding genes were predicted. Evolutionarily accelerated genes related to the phenotypic trait of anti-infective activities for influenza virus were identified. These genes encode three integral membrane proteins, a teichoic acid export ATP-binding protein and a glucosamine - fructose-6-phosphate aminotransferase involved in host innate immunity, the nonspecific DNA-binding protein Dps, which protects bacteria from oxidative damage, and the response regulator of the three-component quorum-sensing regulatory system, which is related to the capacity of adhesion to the surface of the respiratory tract and competition with pathogens. This is the first study to identify the genetic backgrounds of the antiviral activity in L. plantarum strains. These findings provide insight into the anti-infective activities of L. plantarum and the development of preventive probiotics.
Kim, Jong-Chul;Yoon, Jang-Won;Kim, Jong-Bae;Oh, Kyung-Hwan;Park, Mi-Sun;Lee, Bok-Kwon;Cho, Seung-Hak
Journal of Microbiology and Biotechnology
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v.20
no.2
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pp.415-424
/
2010
The purpose of this study was to investigate the relationship between the global regulatory mechanism known as quorum sensing and expression of virulence factors in Escherichia coli O157:87. A nonpolar luxS deletion was introduced into the chromosome of strain CI03J, a human clinical isolate from South Korea, to create the ${\Delta}luxS$ mutant strain ML03J. Phenotypic characterization of wild-type and mutant strains demonstrated that ML03J had no obvious growth or metabolic defects on 0.2% glucose LB medium, produced a functionally defective flagellum, and could not utilize sorbose; the biological significance of sorbose utilization is unknown. Omics-based analysis revealed the involvement of LuxS in the transcriptional activation of several flagella/chemotaxisrelated genes (flhD; fliA, C, D, S, Z; and cheA, Y, Z), repression of glutamate-dependent acid resistance genes (gadAB), and expression of virulence factors including Shiga toxin, hemolysin, and SepD within the LEE pathogenicity island.
KSII Transactions on Internet and Information Systems (TIIS)
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v.7
no.8
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pp.1890-1910
/
2013
Routing overlay offers an ideal methodology to improve the end-to-end communication performance by deriving a backup path for any node pair. This paper focuses on a challenging issue of selecting a proper backup path to bypass the failures on the default path with high probability for any node pair. For existing backup path selection approaches, our trace-driven evaluation results demonstrate that the backup and default paths for any node pair overlap with high probability and hence usually fail simultaneously. Consequently, such approaches fail to derive a robust backup path that can take over in the presence of failure on the default path. In this paper, we propose a three-phase RBPS approach to identify a proper and robust backup path. It utilizes the traceroute probing approach to obtain the fine-grained topology information, and systematically employs the grid quorum system and the Bloom filter to reduce the resulting communication overhead. Two criteria, delay and fault-tolerant ability on average, of the backup path are proposed to evaluate the performance of our RBPS approach. Extensive trace-driven evaluations show that the fault-tolerant ability of the backup path can be improved by about 60%, while the delay gain ratio concentrated at 14% after replacing existing approaches with ours. Consequently, our approach can derive a more robust and available backup path for any node pair than existing approaches. This is more important than finding a backup path with the lowest delay compared to the default path for any node pair.
Recently, quorum sensing has been implicated as an important global regulator controlling the production of numerous virulence factors such as capsular polysaccharides in bacterial pathogens. The nucleotide and deduced amino acid sequences of smcR, a homolog of V. harveyi luxR identified from V. vulnificus ATCC29307, were analyzed. The amino acid sequence of SmcR from V. vulnificus was 72 to 92% similar to those of LuxR homologs from Vibrio spp. Functions of SmcR were assessed by the construction of an isogenic mutant, whose smcR gene was inactivated by allelic exchanges, and by evaluating its phenotype changes in vitro and in mice. The disruption of smcR resulted in a significant alteration in biofilm formation, in type of colony morphology, and in motility. When compared with the wild-type, the smcR mutant exhibited reduced survival under adverse conditions, such as acidic pH and hyperosmotic stress. The smcR mutant exhibited decreased cytotoxic activity toward INT 407 cells in vitro. Furthermore, the intraperitoneal $LD_{50}$ of the smcR mutant was approximately $10^2$ times higher than that of parental wild-type. Therefore, it appears that SmcR is a novel global regulator, controlling numerous genes contributing to the pathogenesis as well as survival of V. vulnificus.
Mechesso, Abraham F.;Quah, Yixian;Park, Seung-Chun
Journal of Ginseng Research
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v.45
no.1
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pp.75-85
/
2021
Background: Invasive infections due to foodborne pathogens, including Salmonella enterica serovar Typhimurium, are prevalent and life-threatening. This study aimed to evaluate the effects of ginsenoside Rg3 (Rg3) on the adhesion, invasion, and intracellular survival of S. Typhimurium. Methods: The impacts of Rg3 on bacterial growth and host cell viability were determined using the time kill and the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assays, respectively. Gentamicin assay and confocal microscopic examination were undertaken to determine the effects of Rg3 on the adhesive and invasive abilities of S. Typhimurium to Caco-2 and RAW 264.7 cells. Quantitative reverse transcription polymerase chain reaction was performed to assess the expression of genes correlated with the adhesion, invasion, and virulence of S. Typhimurium. Results: Subinhibitory concentrations of Rg3 significantly reduced (p < 0.05) the adhesion, invasion, and intracellular survival of S. Typhimurium. Rg3 considerably reduced (p < 0.05) the bacterial motility as well as the release of nitrite from infected macrophages in a concentration-dependent manner. The expression of genes related to the adhesion, invasion, quorum sensing, and virulence of S. Typhimurium including cheY, hilA, OmpD, PrgK, rsgE, SdiA, and SipB was significantly reduced after Rg3 treatment. Besides, the compound downregulated rac-1 and Cdc-42 that are essential for actin remodeling and membrane ruffling, thereby facilitating Salmonella entry into host cells. This report is the first to describe the effects of Rg3 on "trigger" entry mechanism and intracellular survival S. Typhimurium. Conclusion: Rg3 could be considered as a supplement agent to prevent S. Typhimurium infection.
Biofilms significantly affect the performance of wastewater treatment processes in which biodegradability of numerous microorganisms are actively involved, and various technologies have been applied to secure microbial biofilms. Understanding changes in biofilm characteristics by regulating expression of signaling molecules is important to control and regulate biofilms in membrane bioreactor, i.e., biofouling. This study investigated effects of addition of acyl-homoserine lactones (AHL) as a controllable factor for the microbial signaling system on biofilm formation of Pseudomonas aeruginosa PAO1 and multiple strains in membrane bioreactor. The addition of three AHL, i.e., C4-, C6-, and C8-HSL, at a concentration of 200 ㎍/L, enhanced the formation of the PAO1 biofilm and the degree of increases in the biofilm formation of PAO1 were 70.2%, 76.6%, and 72.9%, respectively. The improvement of biofilm formation of individual strains by C4-HSL was an average of 68%, and the microbial consortia increased by approximately 52.1% in the presence of 200 ㎍/L C4-HSL. CLSM images showed that more bacterial cells were present on the membrane surface after the AHL application. In the COMSTAT results, biomass and thickness were increased up to 2.2 times (PAO1) and 1.6 times (multi-strains) by C4-HSL. This study clearly showed that biofilm formation was increased by the application of AHL to individual strain groups, including PAO1 and microbial consortia, and significant increases were observed when 50 or 100 ㎍/L AHL was administered. This suggests that AHL application can improve the biofilm formation of microorganisms, which could yield an enhancement in efficiency of biofilm control, such as in various biofilm reactors including membrane bioreactor and bioflocculent systems in water/wastewater treatment processes.
As the first step to improve the medical recording for the hospitalized patients, we tried to identify problems of completion rate of the medical recording and made the completion rate of the medical recording within due date known to the public in the hospital and commended the best Department and resident officially. The results were as follows: 1. The average number of the medical recordings per a recorder by years of residency(average number to quorum) was 293.3(398.8) in 1st year, 82.5(68.1) in 2nd year, 21.2(8.3) in 3rd year, and 20.5(1.9) in 4th year-residents. There was only 2 residents who prepared the medical recordings more than the average of 125.6 recordings per a resident. 2. Among 13 Departments, the medical recording was wholly put in charge of the 1st year-resident in 6 Departments. The duty was shared with the 2nd year-resident in 5 Departments and the 3rd year-resident in 1 and the 4th year-resident in Only 1 Department. 3. The more the cases requiring the medical recording, the lower the completion rate(80% less than 100 recordings, 70% in 100-299, 60% in 300-399 and 33.3% in more than 400). 4. There was no difference in the completion rate of the medical recording before(1991) and after(1993) a public nitice(1992). However, 4 Departments showed improvement of 21-45% in the completion rate, and no case was found where billing for medical assurance was postponed due to delayed completion of the medical recording. 5. The completion rate was relatively low(72-78%) from January to March. The main reasons were shortage of men power due to preparation of board examination, attendance to military duty, and lack of training in the medical recording for the new 1st year-residents. 6. The official commendation of the best Department and resident by letters did not improve the completion rate of the medical recording. In conclusion, The main reason of the low completion rate of the medical recording was the fact that the 1st year-residents were almost exclusively responsible for the medical recording. Hence, it is mandotory that this practice gets staffs' attention to improve the completion rate. Public notice of the completion rate of the medical recording shows prompt improvement of the completion rate. Prize money rather than commendation by a letter for the best Department would also be more effective to improve the quality of medical recording.
This study is a series of reviews in connection with the application of the Questionnaire THI, in order to intend to provide such datum to be of a help as the screening-test by means of getting through comparative analysis, and finding out existence of any disease appeared as a result of the general medical examinations, and the psychosomatic symptoms, through the questionnaire THI. This study had been researched through the 507 insureds of the life insurance residing in Seoul and in the suburbs of Seoul, during the period 5 Jan. 1987 to 27 Feb. through the questionnaires. Exception of the 88 insincere respondents out of the total objectives, the 419 questionnaires were nanlized through percentage, Mean, T-test, ANOVA test, and Discriminant Analysis. 1. The Subjective Health Problem of the Objectives: The average of the health problem appliation quorum appeared to be 1. 81, the articles of both nervousness and aggressiveness appeared to be each 2.25 and 2.04. It tells that the state of appliation is heavy. It was very significant statistically the relation between the elements of sex, ages, educational backgrounds and occupations. 2. The result of the General Examination of the Objectives: The number of the persons with abnormal findings was 300 persons among the total of 419 persons as a result of the general medical examinations, and it was applicable to $73.5\%$. By each age and marital state were statistically significant. 3. Comparison of the result of the General Examination and Subjective Health of those Objectives: As a result of discriminant analysis of the independent variables through the result of the general examination, the greatest element for influence appeared to be the age, and then, regular examination, sex, and the somatic complaints of the THI in order. Accordingly, it is considered to be able to function as the screening test of the general examination, the THI. In case of a greater unit of group required a medical examination, It is considered to be of a help for the health management service for more effective and precise in quality and economic, only if a medical examination is carried out after selecting the somatic subjective problem and research in advance to the application of the THI prior to the examination.
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