Yoon, Ho Joo;Kim, Mi Ok;Sohn, Jang Won;Kim, Jung Mogg;Shin, Dong Ho;Park, Sung Soo
Tuberculosis and Respiratory Diseases
/
v.54
no.1
/
pp.104-113
/
2003
Background : Rhinovirus(RV) infections frequently trigger dyspnea and paroxysmal cough in adult patients with asthma and are the most prevalent cause of the common cold. However, the mechanisms of a RV-induced airway inflammation is unclear. Since the RV does not directly destroy the airway epithelium, it is presumed that the immune response to the RV contributes to the pathogenesis of the respiratory symptoms. In order to test this hypothesis, this study characterized the time-sequenced alterations in interleukin(IL)-8 elaboration from the human bronchial epithelial cells and evaluated the role of NF(nuclear factor)-${\kappa}B$ in the RV-induced IL-8 production by pretreating the inhibitors of NF-${\kappa}B$ activation. Methods : The ability of RV-infected human bronchial epithelial cells and BEAS-2B cells to produce the IL-8 was compared with the controls. This study infected BEAS-2B cells with the RV14 obtained from the American Type Culture Collection. The supernatants were harvested from the RV infected BEAS-2B cells and the controls at 2hr, 4hr, 6hr, 12hr, 24hr, 48hr from the inoculation time. This study measured the IL-8 concentration using the ELISA kits. In order to elucidate the role of NF-${\kappa}B$ in the RV-induced IL-8 production, the effect of the NF-${\kappa}B$ inhibitors was evaluated on RV-induced IL-8 production. Results: The BEAS-2B cells produced small amounts of IL-8 that accumulated slowly with time in the culture. The RV was a potent stimulator of the IL-8 proteins production by BEAS-2B human bronchial epithelial cells. Antioxidants, N-acetyl-L-cysteine(NAC),\ and pyrrolidine dithiocarbamate(PDTC), blocked the IL-8 elaboration by the RV-infected BEAS-2B cells, which was dose-dependent, but N-Tosyl-L-phenylalanine chloromethyl ketone(TPCK) did not. Conclusion: Some antioxidants inhibited the RV-induced IL-8 production by blocking the NF-${\kappa}B$, which may have a therapeutic potential in asthma.
The objective of this study was carried out to examine the selection effects of in vitro matured porcine follicular oocytes with polar body extrusion and early cleavage as non-invasive marker to know the developmental competence in advance. The porcine oocytes matured for 48 h were examined the polar body extrusion. The examined oocytes were matured for additional $16{\sim}18h$ and activated with 7% ethanol and cultured in $5{\mu}g/ml$ cytochalasin B for 5 h for diploid formation. The treated oocytes were cultured and examined the cleavage after 48 h and continued culturing for 5 days. The oocytes of 21.9% were discarded in morphological selection and 32.1% oocytes were discarded by failure of first polar body extrusion. The selected oocytes were matured and activated and then after 48 h the cleavage rates were examined. In morphologically selected oocytes, 15.8% oocytes were not cleaved and 52.6% oocytes were normally cleaved and 31.6% oocytes were hyper-cleaved over 8-cell stage. However in the first polar body extruded oocytes, 7.1% oocytes were not cleaved and 73.1% oocytes were normally cleaved and 19.8% oocytes were hyper-cleaved. The morphologically selected embryos that not cleavage-selected were developed in 16.7% up to blastocyst and the morphologically selected and cleavage-selected embryos were developed in 31.7%. The polar body extruded oocytes that were not carried out cleavage selection were developed in 39.0% and the polar body extruded and cleavage-selected embryos were developed 49.0%. The first cleavage timing was examined with 12 h interval after activation. In $0{\sim}12,\;12{\sim}24,\;24{\sim}36,\;and\;36{\sim}48h$ intervals, 4.1%, 68.6%, 19.1%, and 2.3% oocytes were cleaved and 5.9% oocytes were not cleaved until 48 after activation. The cleaved oocytes in each interval were cultured and developed upto blastocyst with 0, 39.1, 9.5, and 0%, respectively. This results suggests that polar body extruded and cleaved at $12{\sim}36h$ embryo has higher developmental potential than the others.
Bioactive components involved in the tight junction stabilization of intestinal epithelial cells from Korean red ginseng were studied by analyzing transepithelial electrical resistance (TEER) values of the Caco-2 cell monolayer between the apical and basolateral sides for 96 hr. The treatment with less than $20\;{\mu}g/mL$ of the Korean red ginseng extract to the apical side of Caco-2 cell monolayer gave higher TEER values than the control. However, the treatment with more than $130\;{\mu}g/mL$ of the Korean red ginseng extract drastically decreased the TEER values, and these effects were not due to its cytotoxicity. When fractions of low molecular weight compounds, polysaccharides, proteins, saponins, and polyphenols derived from Korean ginseng were applied to the apical side of the Caco-2 cell monolayer, polyphenols showed high tight junction stabilizing activity and saponins showed low activity, but the others showed no significant activity. These results suggest that Korean red ginseng might be useful for the prevention of food allergy by stabilizing the tight junction of intestinal epithelial cells leading to hindering absorption of food allergens.
This study was carried out to evaluate the effects of condensed molasses soluble (CMS) treatment on the mineralization of N, chemical properties and soil microbial population under the incubation condition with unsaturated water content during the 7 weeks at $25^{\circ}C$ in the different levels of CMS application. The results indicated that the total nitrogen content of soil was increased with increasing application rate of CMS and this trend was maintained up to 7 weeks. With CMS treatment content of $NH_4-N$ was gradually decreased. However, the content of $NO_3-N$ in the soil was gradually increased with incubation time due to the nitrification under the unsturated water condition. The CMS treatment increased the microbial populations such as bacteria, actinomycetes and fungi, which may be due to the availability of more nutrients such as amino acids, sugars and other minor elements from CMS. The pH of soil was found to be reduced by the addition of CMS, Whereas, electrical conduvtivity of soil was correspondingly increased with increasing application rate of CMS.
Five hundred strains of actinomycetes were screened for inhibitory activity against the phytopathogenic fungi; Alternaria alternata, Botryotinia fuckeliana, Colletotrichum acutatum, Colletotrichum gleosporioides, Corticium sasaki, Cylindrocarpon destructans, Fusarium oxysporium, Magnaporthe grisea, Phytophthora infestans, Phytium ultimum, and Thanatephorus cucumeris. The strain CNU-A91231 (Korea Agricultural Culture Collection #20938) showed a strong activity against the phytopathogenic fungi and it was identified as Streptomyces griseofuscus based on the sequence of 16s rDNA. Practical and simple media for the strain S. griseofuscus CNU-A91231 was developed at the conditions ($28^{\circ}C$ and pH 6 with aeration) for efficient bacterial growth. Alanine, glutamine, proline and ammonium ion were good nitrogen sources for the bacterium. Addition of the major salts including Na, Cl, Ca, P, K, and Mg into molasses did not increase the growth of S. griseofuscus. Addition of fertilizers containing amino acids significantly enhanced growth of the bacterium. The optimal medium was formulated as molasses + 1% of glutamate fermentation waste powder. All the conditions and components used in this study did not affect the antifungal activity of S. griseofuscus. The bacterium and the medium in this study can be used as a bio-antifungal agent for plant farming.
To extract insoluble proteins and to improve functional properties of abolished proteins, a protease producing Aspergillus sp. MS-18 was isolated from soil. The enzyme was purified and its enzymological characteristics were investigated. It was found that production of protease reached to the maximum when the wheat brae medium containing, 3% arabinose, 0.5% polypepton, 0.1% $(NH_4)_2SO_4$ and 0.2% magnesium chloride was cultured for 3 days. Protease was purified 16.9 folds after ion exchange chromatography and gel filtration and the specific activity was 340.4 unit/mg. Purified enzyme was confirmed as a single band by the polyacrylamide gel electrophoresis. The molecular weight of protease was estimated to be 30,000. Crystalization form of purified protease was a stick shape with rounding edges. The optimum pH and temperature for the protease activity were 9.0 and $60^{\circ}C$, respectively. The enzyme was stable in pH 7.0-12.0 at $50^{\circ}C$. The activity of purified enzyme was inhibited by $Hg^{2+}$, $Cu^{2+}$, $Zn^{2+}$ and $Pb^{2+}$, whereas it was activited by $Na^+$, $Mg^{2+}$ and $Mn^{2+}$. The activity of the protease was inhibited by the treatment with ethylenediaminetetraacetic acid and phenylmethane sulfonyl fluoride. The result suggests that the purified enzyme is a serine protease with metal ion at active site. Km and Vmax of purified protease were $29.33\;{\mu}mole/L$ and $5.13\;{\mu}g/min$, respectively.
Microorganisms capable of accumulating poly-p-hydroxybutyric acid(PHB) were isolated from soil by enrichment culture technique. Among them, the strain designated as FL-027 had high PHB productivity and was identified as Alcaligenes. The optimal medium composition for cell growth was 8.0 $g/\ell$ of fructose and 3.0 $g/\ell$ of $(NH_4)_2S0_4$, equivalent to C/N ratio 5.04 at pH 7.0 and $30^{\circ}C$. To investigate the optimal conditions for the PHB accumulation, we divided the process into two stages; the first stage for the growth of the cell in nutrient-rich medium and the second stage for the PHB accumulation in nutrient-deficiency medium. The optimal conditions for PHB accumulation were 8.0 $g/\ell$ of fructose and 0.25 $g/\ell$ of $(NH_4)_2S0_4$, equivalent to C/N ratio 60 at pH 6.5 and $30^{\circ}C$. PHB accumulation was stimulated by deficiency of nutrients such as $NH_4^+$, $Ca^{2+}$, $SO_4^{2+}$ in medium. Among them. $NH_4^+$ deficiency was chosen because of its effectiveness. We found the inhibition of cell growth by fructose in batch culture. In order to keep the fructose concentration at an optimal level, intermittent feeding fed-batch culture was employed, and the cell concentration as high as 10.83 $g/\ell$ whose PHB content was responsible for 43% of the dry cell weight. The purified PHB was identified as homopolymer of 3-hydroxybutyric acid by using IR and $^1H-NMR$.
As a basic study for environment-friendly production of bacterial cellulose (BC) dressing with antimicrobial activity, we isolated and identified acetic acid bacteria which are resistant to silver ions and can biosynthesize silver nanoparticles. Furthermore, conditions of BC production by selected strain were also investigated. Strain G7 isolated from decayed grape skin was able to grow in the presence of 0.1 mM $AgNO_3$ which was identified as Acetobacter intermedius based on 16S rRNA gene analysis. BC production was the highest in a medium containing 2% glucose as a carbon source, 2% yeast extract as a nitrogen source, and 0.115% acetic acid as a cosubstrate. Structural properties of BC produced in optimal medium were studied using Fourier-transform infrared spectroscopy and X-ray diffractometer, and it was found that BC produced was cellulose type I that was the same as a typical native cellulose. When strain G7 was cultured in an optimal medium containing 0.1 mM $AgNO_3$, the color of the culture broth turned into reddish brown, indicating that silver nanoparticles were formed. As a result of UV-Vis spectral analysis of the culture, it was found that a unique absorption spectrum of silver nanoparticles at 425 nm was also observed. Scanning electron microscopic observations showed that silver nanoparticles were formed on the surface and pores of BC membrane.
Journal of the Korean Society of Food Science and Nutrition
/
v.25
no.6
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pp.915-921
/
1996
An alkaline pretense Producing microorganism was isolated from Korean traditional soy sauce and identified as Bacillus subtilis CCKS-111. The optimum culture condition of Bacillus subtilis CCKS-111 for the production of alkaline pretense was as follow: 2% soluble starch, 0.2% peptone, 0.1% (NB$_4$)$_2$S$_2$O$_{8}$ , 0.2% MgSO$_4$, pH 7.0, 35$^{\circ}C$ and 24hrs. The optimum pH and temperature for the enzyme activity of alkaline pretense producing Bacillus subtilis CCKS-111 were pH 9.0 and 5$0^{\circ}C$, respectively. The enzyme was relatively stable at pH 6.0~11.0 and at temperature below 4$0^{\circ}C$. The activity of the enzyme was inhibited by $K^{+}$ and Hg$^{2+}$, whereas Cu$^{2+}$ exhibited rather activating effects on the enzyme activity. Ethylenediaminetetraacetic acid and phenylmethanesulfonyl fluoride inhibited the enzyme activity. This indicates that this is serine pretense which requires metal ion group for the enzyme activity. Km value was 2.313$\times$10$^{-4}$ M/L, V$_{max}$ value was 39.216$\mu\textrm{g}$/min. This enzyme hydrolyzed casein more rapidly than the hemoglobin.lobin.
Lee Sang-Sook;Cho Young-Rok;Chun Ji-Min;Choi Yong-Seok;Sohn Eun-Ju;Park Nam-Cho;Park June-Sik
Korean Journal of Head & Neck Oncology
/
v.12
no.2
/
pp.169-176
/
1996
Tuberculous cervical lymphadenitis is still an important cause of neck mass in Korea. Tuberculosis is an important differential diagnosis in patients of cervical lymphadenopathy. Rapid and sensitive test for the diagnosis of tuberculosis is essential for the approapiate treatment. Up to now, conventional diagnostic methods for M. tuberculosis were acid-fast bacilli(AFB) stain and culture of M. tuberculosis. The direct microscopic examination of AFB by Ziehl-Neelsen stain is rapid, but often negative. The culture for M. tuberculosis is time-consuming, taking 4 to 8 weeks. Recently various methods to detect Mycobacterial DNA, including PCR and restriction fragment length polymorphism(RFLP) analysis have been reported. Here we represent a simple method for the confirmation of M. tuberculosis and exclusion of the other Mycobacterial species by RFLP analysis and silver staining of polyacrylamide gel electrophoresis after nested PCR for a repetitive DNA sequence(IS986) specific for M. tuberculosis from fresh or paraffin-embedded biopsy specimens. This result leads us to conclude that this method is simple, rapid and possibly applicable to confirm M. tuberculosis and rule out the other Mycobacteria species from the clinical specimens in the clinical laboratories.
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