• Title/Summary/Keyword: 세포 활성

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Cloning of the β-Lactamase Gene from Bacillus sp. J105 and Analysis of Its Expression in E. colis Cells (Bacillus sp. J105 유래 β-lactamase 유전자의 cloning 및 E. coli 내에서의 발현 분석)

  • Kang, Won-Dae;Lim, Hak-Seo;Seo, Min-Jeong;Kim, Min-Jeong;Lee, Hye-Hyeon;Cho, Kyeong-Soon;Kang, Byoung-Won;Seo, Kwon-Il;Choi, Yung-Hyun;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.18 no.11
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    • pp.1592-1599
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    • 2008
  • The $\beta$-lactamase gene was cloned into E. coli DH5$\alpha$ from Bacillus sp. J105 with strong resistance against $\beta$-lactam antibiotics. The chromosomal DNA was partially digested with Sau3AI and ligated to BamHI digested pLAFR3. $\beta$-Lactamase positive clones were obtained by using in vitro packaging kit. The pKL11-${\Delta}4.6$ with $\beta$-lactamase activity was obtained by subcloning of the recombinant plasmid ($\beta$-lac +). The 6.5 kb fragment in the subcloned plasmid was sequenced. The DNA fragment that contains the $\beta$-lactamase gene encodes 309 amino acids. The 0.17 kb upstream region was similar to those of B. thuringinesis and B. cereus with 97% identity. The deduced amino acids sequence was also similar to those of $\beta$-lactamase from B. thuringinesis and B. cereus with 97% and 94% identity, respectively. The phylogenetic tree also showed the relationships of the $\beta$-lactamase gene of Bacillus sp. J105 to genetically related that of other Bacillus strains. Analysis of expression pattern of the pKL11-${\Delta}4.6$ in E. coli, revealed that the secretion efficiency of $\beta$-lactamase was $4{\sim}5%$ and the molecular weight was as same as that of original $\beta$-lactamase (31 kDa) from Bacillus sp. J105.

Establishment of Reference Value of Insulin Using the Statistical Analysis (통계적 분석을 통한 Insulin의 정상 참고치 설정)

  • Kim, Whe-Jung;Yoon, Pil-Young;Shin, Young-Goon;Yoo, Seon-Hee;Cho, Shee-Man
    • The Korean Journal of Nuclear Medicine Technology
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    • v.14 no.1
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    • pp.143-146
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    • 2010
  • Purpose: Insulin is involved in carbohydrate metabolism and also it's very important because it increases storage of glycogen, synthesis of fatty acids, absorption of amino acid, synthesis of protein. Insulin is clinically useful when we evaluate fasting patients in hypoglycemia, classify and predict diabetes, assess the activity of ${\beta}$-cell, research insulin resistance. We are going to increase usability of insulin assay by establishing normal reference value according to statistical analysis. Material & Method: We selected 6,648 patients who visited asan health medical center from May to August in 2008. We set exclusion criteria as family of diabetes, diabetes medication, the past history of blood glucose rise, more than 100 mg/dL in normal fasting blood glucose, outside the scope of BMI 18.5~22.9 $kg/m^2$, and more than HbA1c 6.5%. We determine whether the subgroup is portioned as sex and age or not and establish normal reference value by conducting statistical analysis as Bayesian's method and Hoffman's method. Result: Portioning of subgroup as sex and age is not needed. By statistical analysis of Bayesian method, results 1.5-11.0 uIU/mL. By statistical analysis of Hoffman method, results 1.8~12.8 uIU/mL. Conclusion: We established 1.8~12.8 uIU/mL as Insulin normal reference value by Hoffman method. This is a similar value with reporting reference value 1.7~11.8 uIU/mL in kit. This will enhance the usability of insulin assay by establishing normal reference value.

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Soluble Interleukin-2 Receptor(sIL-2R) Levels in Patients Tuberculous Pleurisy VS Nontuberculous Pleurisy (결핵성 늑막삼출과 비결핵성 늑막삼출에서의 가용성 Interleukin-2 수용체의 농도)

  • Lim, Hyun-Oak;Ham, Jong-Yeol;Shim, Dae-Seok;Hwang, Young-Sil
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.2
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    • pp.135-143
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    • 1994
  • Background: The cell mediated immunity has an important role in the pathogenesis of tuberculosis. sIL-2R has been known as a sensitive marker of T lymphocyte activation Elevated serum levels of sIL-2R have been found in patients with lymphoproliferative disorders, organ transplantation, autoimmune diseases, and various granulomatous diseases. Elevated levels of sIL-2R have been also found in the serum and pleural fluid of the patients with tuberculosis. To evaluate the diagnostic value of sIL-2R in the differentiation of tuberculous pleurisy and nontuberculous pleurisy. We measured the level of sIL-2R in the sera and pleural fluids of 12 patients with tuberculous pleurisy and 32 patients with nontuberculous pleurisy. Method: Samples of pleural fluid and serum were centrifuged at 2500 rpm for 10 min to remove cell pellets. Soluble IL-2R was measured with a sandwitch enzyme immunoassay using the Cellfree(r) Interleukin-2 Receptor Test kit(T-cell science,Inc. Cambridge, MA). Results: The results obtained were as follows: 1) The sIL-2R level in pleural fluid of the patients with tuberculous pleurisy was higher than that of patients with nontuberculous pleurisy(P<0.005). 2) When the sIL-2R level above 5,000 u/ml in pleural fluid was used as the cut-off value to diagnose tuberculous pleurisy, it had a sensitivity of 84.6% and a specificity of 90.9%. 3) The sIL-2R level in the sera of the patients with tuberculous pleurisy was higher than that of patients with bacterial pleural effusions and normal control group(P<0.05) and there was no difference of levels compared with malignant pleural effusions and transudative pleural effusions(P>0.05). 4) In patients with tuberculous pleurisy, the mean concentration of sIL-2R in pleural fluid was higher than that in serum(P<0.005). Conclusion: These findings suggest that the measurement of elevated levels of pleural fluid sIL-2R in tuberculous pleurisy may be useful in the differential diagnosis between patients with tuberculous pleurisy and nontuberculous pleurisy.

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Development Process for Decreasing Bitterness of Doraji (Platycodon grandiflorum) (도라지 쓴맛 개선을 위한 공정개발 연구)

  • Chang, Yoon-Je;Kim, Eunmi;Choi, Yun-Sang;Jeon, Ki-Hong;Kim, Young-Boong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.10
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    • pp.1550-1557
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    • 2015
  • This study was conducted to evaluate the improvement effects of bitter taste of doraji (Platycodon grandiflorum) with different freezing temperature conditions ($-5^{\circ}C$, $-10^{\circ}C$, $-20^{\circ}C$, and $-70^{\circ}C$). After freezing for 4 h, frozen doraji was thawed and dehydrated. Doraji frozen at $-20^{\circ}C$ showed higher contents of crude fat, crude protein, crude ash, crude fiber, and nitrogen free extract. Moisture, crude saponin, and total polyphenol contents of doraji frozen at $-5^{\circ}C$ were higher than in other samples. Weight loss of doraji was significantly different as freezing temperature decreased and the process repeated. Doraji frozen at $-5^{\circ}C$ showed higher contents of saponin and phenol than the control. Hardness of doraji increased as freezing temperature decreased. According to the preference test, Doraji frozen at $-5^{\circ}C$ showed higher preference and less bitter taste intensity than other samples. These results mean that doraji frozen at $-5^{\circ}C$ with less bitter taste could be utilized widely as a food material.

Naringin Protects Ovalbumin-induced Asthma through the Down-regulation of MMP-9 Activity and GATA-3 Gene (Naringin에 의한 천식치료 효과연구)

  • Lee, Chang-Min;Chang, Jeong-Hyun;Jung, In-Duk;Jeong, Young-Il;Tae, Noh-Kyung;Park, Hee-Ju;Kim, Jong-Suk;Shin, Yong-Kyoo;Park, Sung-Nam;Park, Yeong-Min
    • Journal of Life Science
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    • v.19 no.6
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    • pp.735-743
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    • 2009
  • The common word flavonoids is often used to classify a family of natural compounds, highly abundant in all higher plants, that have received significant therapeutic interest in recent years. Naringin is associated with a reduced risk of heart disease, neurodegenerative disease, cancer and other chronic diseases; however the molecular basis of this effect remains to be elucidated. Thus we attempted to elucidate the anti-allergic effect of Naringin in ovalbumin (OVA)-induced asthma model mice. The OVA-induced mice showed allergic reactions in the airways. These included an increase in the number of eosinophils in bronchoalveolar lavage (BAL) fluid, an increase in inflammatory cell infiltration into the lung around blood vessels and airways, airway luminal narrowing, and the development of airway hyper-responsiveness (AHR). The administration of Naringin before the last airway OVA challenge resulted in a significant inhibition of all asthmatic reactions. Accordingly, this study may provide evidence that Naringin plays a critical role in the amelioration of the pathogenetic process of asthma in mice. These findings provide new insight into the immunopharmacological role of Naringin in terms of its effects on asthma in mice.

Expressional Analysis of Superoxide Dismutase in Olive Flounder (Paralichthys olivaceus) against Viral Hemorrhagic Septicemia Virus Infection (Viral hemorrhagic septicemia virus (VHSV) 감염에 대한 넙치 superoxide dismutase(Of-SOD)의 발현분석)

  • Lee, Young Mee;Kim, Jung-Eun;Noh, Jae Koo;Kim, Hyun Chul;Park, Choul-Ji;Park, Jong-Won;Kim, Kyung-Kil;Lee, Jeong-Ho
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1371-1377
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    • 2014
  • Superoxide dismutase is a family of important antioxidant metalloenzymes and catalyzes the dismutation of toxic superoxide anions into dioxygen and hydrogen peroxide. A recent study identified the partial superoxide dismutase (SOD) gene in olive flounder (Paralichthys olivaceus). The same study reported that it strongly induced benzo[a]pyrene and that it was an indicator of aquatic oxidative stress responses. However, its transcriptional response against viral infection has not been investigated. In the present study, the spatial and temporal expression profiles were analyzed to investigate the function of Of-SOD in the antiviral response. The Of-SOD transcripts were ubiquitously detected at various levels in diverse tissues in a real-time PCR. The expression of Of-SOD was significantly higher in the muscles, liver, and brain but extremely low in the stomach and spleen. Following a VHSV challenge, the expression of Of-SOD increased within 3 h in the kidneys and decreased to the original level 2 days postchallenge. In muscle, liver, and brain, Of-SOD mRNA was similarly up-regulated at 3-6 h postchallenge and then decreased to the basal level. Although the expression pattern and induction time differed slightly depending on the tissue, the transcript of Of-SOD consistently increased in the acute infection response, but the expression was low in the chronic response. The expression of Of-SOD was induced after the VHSV infection, and Of-SOD was probably involved in the immune response against the viral challenge. These results suggest that SOD may play important roles in the immune defense system of P. olivaceus and perhaps contribute to the protective effects against oxidative stress in olive flounder.

Cerebral-perfusion Reserve after Carotid-artery Stenting: Relationship with Power Spectrum of Electroencephalography (경동맥스텐트삽입술 후의 뇌관류예비능: 뇌파파워스펙트럼과의 연관성)

  • Jeong, Da-hye;Jung, Seokwon;Kwak, Byeonggeun;Kim, Young-Soo;Kim, Soo-kyoung;Kwon, Oh-Young
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.2
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    • pp.144-152
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    • 2016
  • Carotid-artery stenosis may reduce cerebral perfusion, and affect cerebral neuronal activities. We examined the question of whether the recovery of cerebral-perfusion reserve after carotid-artery stenting (CAS) can affect the EEG power-spectrum. Nineteen candidates for CAS were initially recruited. Subtraction imaging of single photon emissary computerized tomography (SPECT) and an electroencephalogram (EEG) were taken twice, before and 1 month after CAS. At each time point, the EEGs were recorded before and after injection of acetazolamide (pre-ACZ EEG and post-ACZ EEG). Finally, 7 patients were enrolled after exclusion of incomplete studies. We obtained the spectral ratio (SR) of each hemisphere. SR was defined as the divided value of the power-spectrum sum of fast activities by that of slow activities. The power-spectrum values between hemispheres were compared using the inter-hemispheric index of spectral ratio (IHISR), and we examined the correlation between the power-spectrum and the cerebral-perfusion reserve. Cerebral-perfusion reserve improved after CAS on the stent side in 6 of 7 patients. In 3 patients with unilateral carotid-artery stenosis, CAS increased SR on the pre-ACZ EEGs, and IHISR on the post-ACZ EEGs. The increases of SR and IHISR were concordant with the increment of cerebral-perfusion reserve. In contrast, the results in the other patients with bilateral stenosis showed complex patterns. The SR of pre-ACZ EEGs and IHISR of post-ACZ EEGs may be useful electrophysiological markers for the blood-flow reserve after CAS in patients with unilateral carotid-artery stenosis, but not in those with bilateral stenosis.

Effect of Dietary Supplementation of Plum or Omija on Growth Performance, Blood Biochemical Profiles and Antioxidant Defense System in Broiler Chickens (육계 사료 내 매실 및 오미자 첨가가 성장, 혈액 생화학 성분 및 항산화 작용에 미치는 영향)

  • Ko, Young-Hyun;Moon, Yang-Soo;Sohn, Sea-Hwan;Jung, Chang-Yong;Jang, In-Surk
    • Korean Journal of Poultry Science
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    • v.39 no.2
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    • pp.121-131
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    • 2012
  • To investigate the effect of dietary supplementation of freeze-dried plum (Prunus mume Siebold and Zucc., PMS) or omija meal (Schizandra chinensis Baill.; SCB) on growth performance, organ weights, blood biochemical profiles and antioxidant defense system, a total of 96, 3-day-old male broiler chickens were assigned to three dietary groups: (1) control diet, (2) control diet supplemented with PMS at 0.2%, (3) control diet supplemented with SCB at 0.2%. In vitro antioxidant activity, plum and omija extracts showed a significantly higher radical scavenging activity (RSA). In particular, omija extract showed much higher RSA than plum extract. Dietary addition of plum or omija did not affect body weight, feed intake, feed conversion and the relative weight of digestive organ in birds. Plasma triglyceride significantly (P<0.05) increased in birds fed the diet supplemented with omija compared with those fed control diet without affecting the other blood biochemical components. Furthermore, reduced form of glutathione (GSH) in the liver or muscle significantly (P<0.05) increased in birds fed the diet fortified with plum and omija. However, the specific activities of superoxide dismutase (SOD), glutathione peroxidase (GPX) and glutathione-S-transferase (GST) and MDA (malondealdehyde) in the intestine, liver and muscle were not altered by dietary antioxidant sources. In conclusion, dietary plum and omija resulted in a positive effect on some antioxidant indicators such as increased in vitro RAS in extracts and in vivo GSH level in the liver and muscle without affecting growth performance. Therefore, dietary addition of 0.2% of plum or omija could be applicable as potential antioxidant sources in broiler chick production.

Antioxidant and anti-inflammatory activities of fruiting bodies of Dyctiophora indusiata (흰망태버섯 자실체의 메탄올 및 열수추출물의 항산화 및 항염증 활성)

  • Nguyen, Trung Kien;Shin, Do Bin;Lee, Kyung Rim;Shin, Pyung Gyun;Cheong, Jong Chun;Yoo, Young Bok;Lee, Min Woong;Jin, Ga-Heon;Kim, Hye Young;Im, Kyung Hoan;Lee, Tae Soo
    • Journal of Mushroom
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    • v.11 no.4
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    • pp.269-277
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    • 2013
  • Dictyophora indusiata is an edible mushroom belongs to Family Phallaceae of Phallales, Basidiomycota. The purpose of this study was to investigate the antioxidant and anti-inflammatory activities of methanol and hot water extracts prepared from fruiting bodies of Dictyophora indusiata. Besides measuring of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, a reducing power and a chelating activity on ferrous ions were also measured to evaluate the antioxidant activity for those extracts. To measure the anti-inflammatory activities for the extracts, nitric oxide(NO) production from lipopolysaccharide(LPS) treated RAW 264.7 macrophage cells and carrageenan-induced acute hind paw edema of rats were investigated. The results showed that the extracts have excellent DPPH scavenging and chelating activity on the ferrous ions compared with positive control. The nitric oxide(NO) production in LPS-stimulated RAW 264.7 macrophage cells were decreased as we increased the concentration of the mushroom extracts. Significant reduction of paw edema of rats were observed at 2~6 h after treatment of methanol and hot-water extracts with 50 mg/kg concentration to the rats which are induced acute hind paw edema by carrageenan administration. Therefore, the experimental results suggested that methanol and hot-water extracts of Dictyophora indusiata fruiting bodies might be used for natural sources of antioxidant and anti-inflammatory agents.

Antioxidant, anti-inflammatory and anti-acetylcholinesterase activities of fruiting bodies of Phellinus xeranticus (기와층버섯 자실체의 메탄올 및 열수추출물의 항산화, 항염증 및 아세틸콜린에스테라제 저해 활성)

  • Nguyen, Trung Kien;Shin, Do Bin;Lee, Kyung Rim;Shin, Pyung Gyun;Cheong, Jong Chun;Yoo, Young Bok;Lee, Min Woong;Jin, Ga-Heon;Kim, Hye Young;Im, Kyung Hoan;Lee, Tae Soo
    • Journal of Mushroom
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    • v.11 no.4
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    • pp.278-286
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    • 2013
  • Phellinus xeranticus is an medicinal mushroom belongs to Family Hymenochaetaceae of Polyporales, Basidiomycota. The purpose of this study was to investigate the antioxidant, anti-inflammatory and anti-acetylcholinesterase activities of methanol and hot water extracts prepared from fruiting bodies of Phellinus xeranticus. Besides measuring of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, a reducing power and a chelating activity on ferrous ions were also measured to evaluate the antioxidant activity of the extracts. To measure the anti-inflammatory activities of the extracts, nitric oxide(NO) production from lipopolysaccharide (LPS) stimulated RAW 264.7 macrophage cells and carrageenan-induced acute hind paw edema of rats were investigated. The results showed that the extracts have excellent DPPH scavenging and chelating activity on the ferrous ions compared with positive controls. The nitric oxide (NO) production in LPS-induced RAW 264.7 macrophage cells were decreased as the concentration of the mushroom extracts increased. Significant reduction of paw edema of rats were observed at 2~6 h after treatment of methanol and hot-water extracts with 50 mg/kg concentration to the rats which are induced acute hind paw edema by carrageenan administration. The anti-acetylcholinesterase activity of the methanol extract of the mushroom showed 83.34% inhibition on AcHE which is lower than that of positive control galanthamine. The experimental results suggested that methanol and hot-water extracts of Phellinus xeranticus fruiting bodies might be used for good sources of antioxidant, anti-inflammatory and anti-acetylcholinesterase agents.