• Journal of Microbiology and Biotechnology
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• 제21권5호
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• pp.483-493
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• 2011

An acid phosphatase (HppA) activated by $NH_4Cl$ was purified 192- and 34-fold from the periplasmic and membrane fractions of Helicobacter pylori, respectively. SDS-polyacrylamide gel electrophoresis revealed that HppA from the latter appears to be several kilodaltons larger in molecular mass than from the former by about 24 kDa. Under acidic conditions (pH${\leq}$4.5), the enzyme activity was entirely dependent on the presence of certain mono- and/or divalent metal cations (e.g., $K^+$,$ NH_4{^+}$, and/or $Ni^{2+}$). In particular, $Ni^{2+}$ appeared to lower the enzyme's $K_m$ for the substrates, without changing $V_{max}$. The purified enzyme showed differential specificity against nucleotide substrates with pH; for example, the enzyme hydrolyzed adenosine nucleotides more rapidly at pH 5.5 than at pH 6.0, and vice versa for CTP or TTP. Analyses of the enzyme's N-terminal sequence and of an $HppA^-$ H. pylori mutant revealed that the purified enzyme is identical to rHppA, a cloned H. pylori class C acid phosphatase, and shown to be the sole bacterial 5'-nucleotidase uniquely activated by $NH_4Cl$. In contrast to wild type, $HppA^-$ H. pylori cells grew more slowly. Strikingly, they imported $Mg^{2+}$ at a markedly lowered rate, but assimilated urea rapidly, with a subsequent increase in extracellular pH. Moreover, mutant cells were much more sensitive to extracellular potassium ions, as well as to metronidazole, omeprazole, or thiophenol, with considerably lowered MIC values, than wild-type cells. From these data, we suggest that the role of the acid phosphatase HppA in H. pylori may extend beyond 5'-nucleotidase function to include cation-flux as well as pH regulation on the cell envelope.

• Nutrition Research and Practice
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• 제3권3호
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• pp.180-184
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• 2009

The apoptotic effect of bacteria-derived $\beta$-glucan was investigated in human colon cancer cells SNU-C4 using terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, reverse transcription-polymerase chain reaction (RT-PCR) expressions of Bcl-2, Bax, and Caspase-3 genes, and assay of caspase-3 enzyme activity. $\beta$-Glucan of 10, 50, and $100{\mu}g$/mL decreased cell viability in a dose-dependent manner with typical apoptotic characteristics, such as morphological changes of chromatin condensation and apoptotic body formation from TUNEL assay. In addition, $\beta$-glucan ($100{\mu}g$/mL) decreased the expression of Bc1-2 by 0.6 times, whereas the expression of Bax and Caspase-3 were increased by 3.1 and 2.3 times, respectively, compared to untreated control group. Furthermore, the caspase-3 activity in the $\beta$-glucan-treated group was significantly increased compared to those in control group (P < 0.05). Bacterial derived $\beta$-glucan could be used as an effective compound inducing apoptosis in human colon cancer.

• 생약학회지
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• 제47권2호
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• pp.150-157
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• 2016

Earth worm (Eisenia andrei) and clam worm (Perinereis linea) have been used as anti-bacterial and anti-inflammatory agent. However, it is unclear how they exerted their physiological effects in macrophages. In this experiment, the anti-inflammatory and antioxidant effects of clam worm extract (CWE) and earth worm extract (EWE) in RAW264.7 cells were examined by measuring MDA, catalase, SOD, GSH-Px and inflammatory cytokines (nitric oxide, iNOS, interleukin-$1{\beta}$ and tumor necrosis factor-${\alpha}$). Treatment with CWE significantly increased the activities of catalase, SOD and GSH-Px in RAW264.7 cells and decreased the level of MDA. Interestingly, treatment with CWE induced more activities of SOD than EWE. In addition, CWE decreased NO production, iNOS, COX-2, TNF-${\alpha}$ and IL-$1{\beta}$ in RAW264.7. The EWE also decreased NO production and iNOS, but increased COX-2 and IL-$1{\beta}$ suggesting that CWE could be better resources for anti-inflammatory and antioxidant agent than EWE. Taken together, these results indicate that CWE has the potential as a natural antioxidant and a therapeutic for inflammation-related diseases.

• 한국산업보건학회지
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• 제1권2호
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• pp.136-143
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• 1991

Inorganic mercury in urine and airborne was determined by cold vapor atomic absorption spectrophotometry. Detailed sampling methods and analylical results are as follows : 1. 100~200ml of urine for each person was taken in 250 ml borosilicate bottle and $K_2S_2O_8$ (0.1g/100ml urine) was added to prevent bacterial contamination. About 1001 air of workingplace was absorbed in l0ml of absorbing solution. Urine samples and absorbing solution tubes were stored at $4^{\circ}C$. Dillution solution to prepare standard solution used deionized water (D.W) for urine and absorbing solution (A.S) for air. 2. 1n this procedure deteclion limit was 1ng/ml and mercury contents of blank reagent solution was 1~2ng/ml. 3. Calibration range was $0.02{\sim}0.1{\mu}g/ml$ and in this range r.s.d for each calibration curve in D.W and A.S and ${\pm}7.9%$ and ${\pm}3.7%$, respectively. 4. Repeatability (n=5 times, conc. $0.05{\mu}g/ml$) was ${\pm}5.8%$, in D.W. and ${\pm}4.4%$ in A.S, respectively. 5. Recovery for urine adding spiked concentration ($0.05{\mu}g/ml$) was about 90%. 6. Analytical result of samples was $1{\sim}139{\mu}g/l$ in urine and ${\sim}0.127mg/m^3$ in airborne.

• 대한한방내과학회지
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• 제29권3호
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• pp.800-809
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• 2008

Objective : Although chronic prostatitis/chronic pelvic pain syndrcme(CP/CPPS) is a common disease, there is no consensus on the etiology or pathology and treatment. This was a double-blinded, placebo-controlled, randomized clinical trial, investigating the therapeutic effects of the traditional Korean medicine, Bosingunyang-tang(BSGYT). Method : Participants who met US National Institutes of Health (NIH) consensus criteria for CP/CPPS were entered after applying inclusion/exclusion criteria. They were randomized to the BSGYT or placebo group. and treated three times a day for 6 weeks. NIH-Chronic Prostatitis Symptom Index (NIH-CPSI) was used to estimate the clinical symptoms of CP/CPPS. Prostaglandin E2 and ${\beta}$-endorphin in prostatic fluid, collected by 2-glass pre-massage and post-massage test, were analyzed as factors associated with pain and inflammation. Result : The mean decrease in NIH-CPSI total score of the BSGYT group was 11.0 points, which is 5.7 points more than the placebo group. (Mann Whitney test P=0.038) Also the BSGYT group showed three times higher response rate than the placebo group in NIH-CPSI pain subscale score. (Fisher's exact test P=0.027) In those responders, prostaglandin E2 decreased significantly. (Wilcoxon's signed-ranks test P=0.037). No specific side effects were observed. Conclusion : After a 6-week treatment period, BSGYT improved clinical symptoms of CP/CPPS patients by decreasing PGE2 level in prostatic fluid.

• Toxicological Research
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• 제24권4호
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• pp.321-328
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• 2008

Smoke flavors based on the thermal decomposition of wood have been applied to a variety of food products as an alternative for traditional smoking. Despite its increasing use, the available genotoxicity data on wood smoke flavors (WSF) are still controversial. Thus, potential genotoxic effects of WSF in four short-term in vitro genotoxicity assays were investigated, which included the Ames assay, chromosomal aberration assay, micronucleus test and the alkaline comet assay. WSF did not cause any mutation in the Ames assay using five tester strains at six concentrations of 0.16, 0.31, 0.63, 1.25, 2.5 and 5 ${\mu}l/plate$. To assess clastogenic effect, the in vitro chromosomal aberration assay was performed using Chinese hamster lung cells. No statistically significant increase in the number of metaphases with structural aberrations was observed at the concentrations of 1.25, 2.5, and 5 ${\mu}l/ml$. The in vitro comet assay and micronucleus test results obtained on L5178Y cells also revealed that WSF has no genotoxicity potential, although there was a marginal increase in micronuclei frequencies and DNA damage in the respective micronucleus and comet assays. Taken together, based on the results obtained from these four in vitro studies, it is concluded that WSF is not a mutagenic agent in bacterial cells and causes no chromosomal and DNA damage in mammalian cells in vitro.

• Molecular & Cellular Toxicology
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• 제5권3호
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• pp.257-264
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• 2009

Gardenia yellow, extracted from gardenia fruit, has been widely used as a coloring agent for foods, and thus, safety of its usage is of prime importance. In the current study, short-term genotoxicity assays were conducted to evaluate the potential genotoxic effects of gardenia yellow. The gardenia yellow used was found to contain 0.057 mg/g of genipin, a known biologically active compound of the gardenia fruit extract. Ames test did not reveal any positive results. No clastogenicity was detected by a chromosomal aberration test, even on evaluation at the highest feasible concentration of gardenia yellow. Gardenia yellow was also shown to be non-genotoxic using an in vitro comet assay and a micronucleus test with L5178Y cells, although a marginal increase in DNA damage and micronuclei frequency was reported in the respective assays. Additionally, in vivo micronucleus test results clearly demonstrated that oral administration of gardenia yellow did not induce micronuclei formation in the bone marrow cells of male ICR mice. Taken together, our results indicate that gardenia yellow is not mutagenic to bacterial cells, and that it does not cause chromosomal damage in mammalian cells, either in vitro or in vivo.

• 한국식품영양학회지
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• 제26권4호
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• pp.766-771
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• 2013

Tomato sauce is widely known as a health food, In this study perilla leaf was added to a tomato sauce and the resulting functional improvements were assessed. The moisture of the tomato sauce WITH 2% added perilla leaf was 89.52%, the highest observed likewise, sample pH also significantly increased with 2% perilla leaf, to the highest pH of 4.71, (p>0.001). Lightness L value of the control group was the highest at 40.70, and the redness a value and yellowness b value were highest in, the control group, at 15.77, and 17.70, respectively. The highest salinity was foung with 2% perilla leaf, 0.96, and the sugar content with 2% perilla leaf, was 9.57. The reducing sugar was highest in the control with 63.08. The change in the total bacterial counts was measured during 9 days of storage at $4^{\circ}C$, When perilla leaf was added to, the tomato sauce, no microorganisms were detected until 0~1 days of storage. After five days of storage, microorganisms were detected in the group with 2% perilla leaf. The control group ($1.8{\times}10^5$ CFU/ml) had the highest levels of microorganisms. As the amount of perilla leaf, the total viable count of microorganisms detected was less. In the acceptability test results, the color 6.9 of the 1.5% treatment group, received the highest evaluation. Flavor and taste of the 2% perilla leaf treatment group were highly appreciated at 6.1 and 6.6, respectively. The ranking of overall acceptability was 1.5%>1%>0.5%>10%>2% of perilla leaf; 2% perilla leaf was the most underrated. Based on these results, up to 1.5% perilla leaf could be added to tomato sauce and the commercial potential could be very high.

• Asian-Australasian Journal of Animal Sciences
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• 제11권6호
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• pp.661-672
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• 1998

Two experiments were conducted to examine the effects of a range of concentrations of ruminal fluid ammonia ($NH_3$-N) on forage digestibility, microbial growth efficiency and the mix of microbial species. Urea was either continuously infused directly into the rumen of sheep fed 33.3 glh of oaten chaff (Exp. I) or sprayed onto the oaten chaff (750 g/d) given once daily (Exp. 2). Concentrations of $NH_3$-N increased with incremental addition of urea (p < 0.01). Volatile fatty acids (VFA) concentrations and 24 h in sacco organic matter digestibility in the rumen were higher when supplemental urea was given (p < 0.01). The (C2 + C4) : C3 VFA ratio was lower (p < 0.05) when $NH_3$-N was above 200 mgN/I. The fungal sporangia appearing on oat leaf blades were significantly higher when urea was supplemented, indicating that $NH_3$-N was a growthlimiting nutrient for fungi at levels of $NH_3$-N below 30 mgN/l. The density of protozoa was highest when $NH_3$-N concentrations were adjusted to 30 mgN/I for continuously fed ($4.4{\times}10^5/ml$) and to 168 mgN/1 for once daily feeding ($2.9{\times}10^5/ml$). Thereafter increasing concentrations of $NH_3$-N, were associated with a concomitant decline in protozoal densities. At the concentration of $NH_3$-N above 200 mgN/l, the density of protozoa was similar to the density of protozoa in ruminal fluid of the control sheep ($1.8{\times}10^5/ml$). The efficiency of net microbial protein synthesis in the rumen calculated from purine excretion was 17-47% higher when the level of $NH_3$-N was above 200 mgN/1. The possibilities are that 1) there is less bacterial cell lysis in the rumen because of the concomitant decrease in the protozoal pool and/or 2) microbial growth per se in the rumen is more efficient with increasing $NH_3$-N concentrations.

• Restorative Dentistry and Endodontics
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• 제20권1호
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• pp.71-95
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• 1995

Bacteria have been regarded as a one of major etiologic factors in root canal infections. In endodontic treatment the effective removal of pathogenic microorganisms in the root canal is the key to successful outcome. Bacterial cell wall components may play an important role in the development of pulpal and periapical disease. The purpose of this study was to evaluate the effect of sonic extracts of Streptococcus spp. on cultured L929 cells and to characterize cell wall protein profiles of Streptococcus spp. Streptococcus spp. were isolated from infected root canals and identified with Vitek Systems(Biomeriux, USA). Five streptococci, namely S. sanguis, S. mitis, S uberis, S. mutans (ATCC 10449) and S. faecalis (ATCC 19433) weere enriched in brain heart infusion broth. Cell pellets were sonicated and cell wall extracts were dialyzed and membrane filtered. Prepared cell wall proteins were applied to cultured L929 cell. The cell reaction were evaluated by monitoring DNA synthesis, cell numbers and the change of cell morphology. The total cell wall protein profiles of microorganisms were characterized by sodium dodecyl sulfate polyacrylamide-gel eledruphoresis(SDS-PAGE). DNA synthesis of L929 cells were reduced by the increasing concentration of sonic extracts. DNA synthesis was significantly suppressed in more than $50{\mu}g$/ml of sonic extract conentration in five streptococci. S. nutans (ATCC 10449) showed stronger suppression on DNA synthesis than remaining four streptococci, which had the similar effect on DNA synthesis. Analysis of DNA synthesis measured by [$^3H$]-thymidine uptake was more sensitvie method than cell counting. Sonic extracts affected the microscopic findings of L929 cells. The protein profiles indicated that all five strains shared two major proteins with molecular masses of 70.8 and 57.5 kD respectively. S. uberis and S. mutans shared common minor proteins of which molecular weights were 147.9 and 112.2 kD respectively. However some minor proteins were unique for S. mitis, S. uberis and S. faecalis.