Development of expression vectors is important for the basic and applied researches on kimchi LAB (lactic acid bacteria). An expression vector, pSJE6c was constructed by inserting P6C promoter sequence from Lactococcus lactis into pSJE, a shuttle vector for E. coli and Leuconostoc species. To test the efficiency of pSJE6c, aga ($\alpha$-galactosidase) and lacZ ($\beta$-galactosidase) genes were expressed in Lactobacillus brevis 2.14. Compared to the pSJE, expression levels of both genes were increased, indicating P6C promoter was better than indigenous promoters. Enzyme activities of L. brevis cells harboring pSJE6caga (pSJE6c with aga) or pSJE6Z (pSJE6c with lacZ) were 1.5-2 fold higher than those with pSJEaga (pSJE with aga) or pSJEZ (pSJE with lacZ). More RNA transcripts were detected in cells harboring pSJE6c based recombinant plasmid. The results indicated that heterologous gene expressions in kimchi LAB could be improved significantly by use of efficient expression vectors.
Kim, Hye Jin;Shin, Kyeong Cheol;Lee, Jae Woong;Kim, Kyu Jin;Hong, Yeong Hoon;Chung, Jin Hong;Lee, Kwan Ho
Tuberculosis and Respiratory Diseases
/
v.59
no.6
/
pp.625-630
/
2005
Background : Determining the cause of an exudative pleural effusion is sometimes quite difficult, especially between malignant and tuberculous effusions. Twenty percent of effusions remain undiagnosed even after a complete diagnostic evaluation, including pleural biopsy. The activity of tumor necrosis factor-alpha (TNF-${\alpha}$), which is the one of proinflammatory cytokines, is increased in both infectious and malignant effusions. The aim of this study was to investigate the diagnostic efficiency of TNF-${\alpha}$ activity in distinguishing tuberculous from malignant effusions. Methods : 46 patients (13 with malignant pleural effusion, 33 with tuberculous pleural effusion) with exudative pleurisy were included. TNF-${\alpha}$ concentrations were measured in the pleural fluid and serum samples using an enzyme-linked immunosorbent assay (ELISA). In addition, TNF-${\alpha}$ ratio (pleural fluid TNF-${\alpha}$ : serum TNF-${\alpha}$) was calculated. Results : TNF-${\alpha}$ concentration and TNF-${\alpha}$ ratio in the pleural fluid were significantly higher in the tuberculous effusions than in the malignant effusions (p<0.05). However, the serum levels of TNF-${\alpha}$ in the malignant and tuberculous pleural effusions were similar (p>0.05). The cut off points for the pleural fluid TNF-${\alpha}$ level and TNF-${\alpha}$ ratio were found to be 136.4 pg/mL and 6.4, respectively. The sensitivity, specificity and area under the curve were 81%, 80% and 0.82 for the pleural fluid TNF-${\alpha}$ level (p<0.005) and 76%, 70% and 0.72 for the TNF-${\alpha}$ ratio (p<0.05). Conclusion : We conclude that pleural fluid TNF-${\alpha}$ level and TNF-${\alpha}$ ratio can distinguish a malignant pleural effusion from a tuberculous effusion, and can be additional markers in a differential diagnosis of tuberculous and malignant pleural effusion. The level of TNF-${\alpha}$ in the pleural fluid could be a more efficient marker than the TNF-${\alpha}$ ratio.
Journal of the Korean Society of Food Science and Nutrition
/
v.39
no.2
/
pp.227-236
/
2010
This study was carried out to investigate the anti-oxidative and anti-inflammatory actions of Hericium erinaceus mycelia in BALB/C mice injected with lopopolysaccharide (LPS), called endotoxin. Mice (6 weeks of age) weighing approximately $24.73\pm0.11$ g were divided into 5 groups and were fed on the experimental diets containing Hericium erinaceus mycelia powder (HMP) for 1 week. Experimental groups were NC (normal control), HMP-C (HMP control), LC (LPS control), HMP 3%, and HMP 10%. Endotoxin shock was induced by intraperitoneal injection of LPS (100 mg/kg BW). NC and HMP-C groups were injected with saline solution (100 mg/kg BW). Food efficiency ratio were significantly (p<0.05) decreased in the HMP supplementation groups. Total fat and $\beta$-glucan excretion were higher in HMP supplementation groups than NC and LC groups, while plasma TG level was not different among groups. Plasma ALT levels were significantly (p<0.05) lower in the HMP supplementation groups than in LC group at 8 hr after LPS injection, while tumor necrosis factor-$\alpha$ and interleukine-6 levels of plasma were not different among groups. Hepatic superoxide dismutase, glutathione-reductase (GSH-red), and glutathione-peroxidase activities were higher in the HMP supplementation groups than in LC group at 4 hr after intraperitoneal injection of LPS. Hepatic GSH levels and protein expression of GSH-red was significantly (p<0.05) higher in the HMP supplemented groups than in LC group at 1 hr, 4 hr and 8 hr after LPS injection. From the above results, it is concluded that Hericium erinaceus mycelia may ameliorate hepatic oxidative stress by LPS through the elevation of hepatic glutathione level and antioxidant enzyme activities, which support the hepatoprotective effect of Hericium erinaceus mycelia.
Journal of the Korean Society of Food Science and Nutrition
/
v.39
no.2
/
pp.219-226
/
2010
The objective of this study was to determine the effects of a Scutellaria baicalensis water extract (SDWE) on lipid levels, lipid peroxidation and antioxidant enzyme activities on rats fed a high fat diet for 6 weeks. Thirty-two male Sprague-Dawley rats (4-weeks-old) were randomly divided into four groups: normal diet and deionized water (ND), normal diet and Scutellaria baicalensis water extract (NDS), high fat diet and deionized water (HFD), high fat diet and Scutellaria baicalensis water extract (HFDS). The food intakes were significantly lower, but the food efficiency ratios were significantly higher in the high fat diet groups than those in other groups. The level of HDL-cholesterol and HDL-cholesterol/total cholesterol ratio in plasma were significantly higher and AI (atherogenic index) in HFDS group was significantly lower than that in HFD group. The level of triglyceride in plasma was significantly decreased in SDWE groups. The triglyceride of liver was significantly increased in the high fat diet groups and the total cholesterol of liver in the HFDS group was significantly lower than that in the HFD group. The plasma and liver concentrations of thiobarbituric acid reactive substances (TBARS) in the NDS group were significantly lower than those in the ND group. The total antioxidant status (TAS) in plasma was significantly increased in the HFDS group compared to the HFD group. The activities of SOD, catalase and GST were significantly increased in SDWE groups compared to ionized water groups. The activity of GSH-Px and the concentration of GSH in liver in the HFDS group were significantly higher than those in the HFD group. These results suggest that a supplement of SDWE on rats fed high fat diet reduce levels of lipid and lipid peroxidation in plasma and liver and improve the antioxidant defense systems.
Gil, Min;Kwon, Hyuck Hwan;Kwon, Young Hyun;Jung, Mi Jin;Kim, Sang Yong;Rhie, Yong Ha
Journal of Bio-Environment Control
/
v.29
no.4
/
pp.344-353
/
2020
Plants native in Korea have not only ornamental values but also have excellent environmental adaptability, so they can be used as garden plants. Studies on proper volumetric water content (VWC) of substrates have been reported, but many have been conducted in glasshouse conditions where environmental factors were controlled. When considering garden planting, it is necessary to perform the automated irrigation system in outdoor conditions where rainfall occurs at frequent intervals. This research aimed to investigate the VWC suitable for the growth of Minuartia laricina, Arenaria juncea, and Corydalis speciosa in open filed. Sandy soil which consisted of particles of weathered rock was used, and the VWC of 0.15, 0.20, 0.25, and 0.30 ㎥·m-3 was maintained using an automated irrigation system with capacitance soil moisture sensors and a data logger. No significant differences in growth and antioxidant enzymes activity of A. juncea were observed among VWC treatments. However, the survival rate was low at VWC 0.30 ㎥·m-3 treatment, which was the highest soil moisture content. Even considering the efficiency of water use, we recommended that VWC 0.15-0.20 ㎥·m-3 is suitable for the cultivation of A. juncea. Minuartia laricina showed better growth with lower VWC. Because of frequent rainfall in open field, plant volume and survival rate was high even in VWC 0.15 ㎥·m-3 treatment. In C. speciosa, the plant height, number of shoots and lateral shoots, and fresh and dry weight were higher in plants grown in VWC 0.25 ㎥·m-3 as compared with that in the plants grown at 0.15, 0.20, and 0.30 ㎥·m-3. Based on these results, M. laricina needed less water in open filed, and A. juncea and C. speciosa required higher VWC, but excessive water should be avoided.
In this study, we investigated the preventive effects of the mulberry leaf tea fermented by Monascus pilosus on high fat-induced obesity, hyperlipidemia, and fatty liver in mice. Non-fermented mulberry leaf tea powder (UM) and fermented mulberry leaf tea powder (FM) were supplemented with high-fat diet at 2% (wt/wt) dosage for 8 weeks. Both UM and FM lowered body weight gain, feed efficiency ratio, epididymal fat, serum triglyceride, total cholesterol and LDL-cholesterol increased markedly with high fat diet (HC) in mice. FM showed more significant effects when it was compared with UM. In addition, Hepatic lipid peroxides and xanthin oxidase activities of the UM and FM were significantly lower than those of HC, despite the lack of a big difference in the amount of hepatic GSH. Activities of ROS scavenging enzymes and serum alanine aminotransferase activity were also examined as a parameter of hepatic damage. The UM and FM groups showed a recovery to NC group from significant changes induced by HC. Finally, histopathological analyses of liver samples revealed a decrease of lipid accumulation in hepatocytes in the UM and FM groups. These results suggest that UM and especially FM can reduce the development of obesity, hyperlipidemia and fatty liver.
This study investigated the hepatoprotective effects of an ethanol extract of lotus root (LRE) on alcohol-induced liver damage in rat. Sprague-Dawley rae weighing $100{\sim}150g$, were divided into 6 groups: basal diet group (BD), alcohol (35% 10 mL/kg/day) teated stoup (ET), LRE 200 mg/kg/day teated group (BD-LREL). LRE 400 mg/kg/day treated group (BD-LREH), LRE 200 mg/kg/day and alcohol treated group (ET-LREL), and LRE 400 3mg/kg/day and alcohol teated group (ET-LREH). After the administration, rats were sacrificed to get serum and liver to analyze antioxidant enzyme activity, glutathione and lipid peroxide contents. The body weight gain and feed efficiency ratio were decreased by alcohol administration, however, were gradually increased to a little lower level than the basal diet group by the combined administration of alcohol and LRE. The serum alanine aminotransferase (ALT), asparate aminotransferase (AST) and alkaline phosphatase (ALP) activities that were elevated by alcohol were significantly decreased by LRE administration. It was also observed that thiobarbituric acid reactive substances (TBARS) content, xanthine oxidase (XO), superoxide dismutase (SOD), catalase and glutathione peroxidase (GSH-Px) activities in liver that were increased by alcohol, were markedly decreased in the combined alcohol and LRE administered groups as compared with the alcohol administrated group. These effect of LRE within the alcohol groups were in a dose-dependent manner. The glutathione (GSH) content in liver was decreased by alcohol administration, however, increased after administering LRE. Teken together, these result suggest that ethanol extract of lotus root may have a possible protective effect on liver function in hepatotoxicity-induced rat by alcohol administration.
Background : Tumor necrosis factor(TNF) has been considered as an important candidate for cancer gene therapy based on its potent anti-tumor activity. However, since the efficiency of current techniques of gene transfer is not satisfactory, the majority of current protocols is aiming the in vitro gene transfer to cancer cells and re-introducing genetically modified cancer cells to host. In the previous study, it was shown that TNF-sensitive cancer cells transfected with TNF-$\alpha$ cDNA would become highly resistant to TNF, and the probability was shown that the acquired resistance to TNF might be associated with synthesis of some protective protein. Understanding the mechanisms of TNF -resistance in TNF-$\alpha$ cDNA transfected cancer cells would be. an important step for improving the efficacy of cancer gene therapy as well as for better understandings of tumor biology. This study was designed to evaluate the role of MnSOD, an antioxidant enzyme, in the acquired resistance to TNF of TNF-$\alpha$ cDN A transfected cancer cells. Method : We transfected TNF-$\alpha$ c-DNA to WEHI164(murine fibrosarcoma cell line), NCI-H2058(human mesothelioma cell line), A549(human non-small cell lung cancer cell line), ME180(human cervix cancer cell line) cells using retroviral vector(pLT12SN(TNF)) and confirm the expression of TNF with PCR, ELISA, MIT assay. Then we determined the TNF resistance of TNF-$\alpha$ cDNA transfected cells(WEHI164-TNF, NCIH2058-TNF, A549-TNF, ME180-TNF) and the changes of MnSOD mRNA expressions with Northern blot analysis. Results : The MnSOD mRNA expressions of parental cells and genetically modified cells of WEHI164 and ME180 cells(both are naturally TNF sensitive) were not significantly different The MnSOD mRNA expressions of genetically modified cells of NCI-H2058 and A549(both are naturally TNF resistant) were higher than those of the parental cells, while those of parental cells with exogenous TNF were also elevated. Conclusion : The acquired resistance to TNF after TNF-$\alpha$ cDNA transfection may not be associated with the change in the MnSOD expression, but the difference in natural TNF sensitivity of each cell may be associated with the level of the MnSOD expression.
To investigate antioxidative effects of phenolic compounds separated from persimmon leaves(PL)(Diospyros kaki Thunb.) on the ethanol-induced hepatotoxicity in rat, Sprague-Dawley rats weighing 100-150 g were divided into 5 groups; control group(CON), PL(70 mg/kg) administered group(PEl), ethanol(5 mL/kg, 25%) administered group(ETH), PL(70 mg/kg) and ethanol administered group (PE2), and PL(140 mg/kg) and ethanol administered group(PE3), respectively. The antioxidative activity of persimmon leaves decreased in order of ethylacetate>interphase materials>n-butanol>chloroform>n-hexane>water fraction. The growth rate and feed efficiency ratio decreased by ethanol were gradually increased to the adjacent level of CON by administering PL. The serum activities of ALT, alkaline phosphatase and lactic acid dehydrogenase elevated by ethanol were decreased significantly. It was also observed that the activities of SOD, catalase, and GSH-Px of rat liver increased by ethanol were markedly decreased in PL administered group as compared to ETH. The GSH content of liver was decreased by ethanol, but that was increased in PE1 and PE2 compared with ETH as a dose-dependant manner. These results suggested that phenolic compounds separated from persimmon leaves have a possible protective and relievable effect on the ethanol-induced hepatotoxicity in rats.
Apple (Malus pumila) is one of the most economically important fruits in Korea. but virus infection has decreased the sustainable production of apples and caused serious problems such as yield loss and poor fruit quality. Virus or viroid infection including apple chlorotic leaf spot virus (ACLSV), apple stem pitting virus (ASPV), apple stem grooving virus (ASGV), apple mosaic virus (ApMV) and apple scar skin viroid (ASSVd) have been also reported in Korea. In many cases, as apple gets infected with virus and viroid with no specific symptoms, the damage and symptoms caused by the viruses are not detected. In our research, viruses in the rootstock were eliminated for a virus-free apple dwarfing rootstock of M.9 and M.26. The virus elimination methods were apical meristem culture, thermotherapy ($37^{\circ}C$, 6 weeks) and chemotherapy($Ribavirin^{(R)}$). The detection of apple viruses was accomplished by Enzyme-linked Immuno-Sorbent Assay (ELlSA) and reverse transcription-polymerase chain reaction (RT-PCR). RT- PCR method was 10 ~ 30% more sensitive than the ELISA method. The efficiency of virus elimination was enhanced in apical meristem culture method. The acquisition rate of virus-free apple dwarfing rootstocks was 30 ~ 40% higher in apical meristem culture. After the meristem culturing of M.9, the infection ratio of ACLSV, ASPV and ASGV was 45%, 60% and 50%, respectively. In the apple dwarfing rootstock of M.26, the infection ratio of ACLSV, ASPV and ASGV was 40%, 55% and 55%, respectively. Based on this study, the best method for the production of virus-free apple dwarfing rootstocks was the apical meristem culture.
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