Park, Ha Ju;Han, Se Jong;Yim, Joung Han;Kim, Dockyu
Korean Journal of Microbiology
/
v.54
no.1
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pp.60-68
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2018
A cold-active and alkaline serine protease (Pro21717) was partially purified from the Antarctic marine bacterium Pseudoalteromonas arctica PAMC 21717. On a zymogram gel containing skim milk, Pro21717 produced two distinct clear-zones of approximately 37 kDa (low intensity) and 74 kDa (high intensity). These were found to have identical N-terminal sequences, suggesting they arose from an identical precursor and that the 37 kDa protease might homodimerize to the more active 74 kDa form of the protein. Pro21717 displayed proteolytic activity at $0-40^{\circ}C$ (optimal temperature of $40^{\circ}C$) and maintained this activity at pH 5.0-10.0 (optimal pH of 9.0). Notably, relative activities of 30% and 45% were observed at $0^{\circ}C$ and $10^{\circ}C$, respectively, in comparison to the 100% activity observed at $40^{\circ}C$, and this enzyme showed a broad substrate range against synthetic peptides with a preference for proline in the cleavage reaction. Pro21717 activity was enhanced by $Cu^{2+}$ and remained stable in the presence of detergent surfactants (linear alkylbenzene sulfonate and sodium dodecyl sulfate) and other chemical components ($Na_2SO_4$ and metal ions, such as $Ba^{2+}$, $Mg^{2+}$, $Ca^{2+}$, $Zn^{2+}$, $Fe^{2+}$, $K^+$, and $Na^{2+}$), which are often included in commercial detergent formulations. These data indicate that the psychrophilic Pro21717 has properties comparable to the well-characterized mesophilic subtilisin Carlsberg, which is commercially produced by Novozymes as the trademark Alcalase. Thus it has the potential to be used as a new additive enzyme in laundry detergents that must work well in cold tap water below $15^{\circ}C$.
Purpose: The purpose of the study was to investigate whether adherence to the Korea Healthy Eating Index (KHEI) was associated with metabolic syndrome and risk markers. Methods: The participants included 8,345 adults, aged 20-59 years, who took part in the 7th Korea National Health and Nutrition Examination Survey (KNHANES). The data were analyzed using a complex-sample t-test, the Rao Scott χ2-test, and logistic regression analysis on the SPSS v. 26.0 software. The participants were divided into four groups by quartiles of KHEI scores. Results: The average KHEI score was 61.06 points out of 100, and the women's score (62.50 points) was significantly higher than that of men (59.63 points). The KHEI quartiles status showed significant differences by age (p < 0.001), household income (p < 0.001), smoking status (p < 0.001), and food security. Specifically, the KHEI quartiles in the men showed significant differences in education (p < 0.001) and economic activity (p < 0.001) whereas those of women showed significant differences in alcohol-consumption (p < 0.001), depression (p < 0.01) and eating-out (p < 0.001). As the KHEI scores increased, the proportion of subjects with an energy intake below the estimated energy requirement (EER) was lower, and significantly better levels of intake were observed for carbohydrate, protein, vitamin C, calcium, vitamin B1, vitamin B2, and niacin. The incidence of the metabolic syndrome risk factors, hypertriglyceridemia and hyperglycemia for men and hypertension, and hyperglycemia for women showed significant differences. The KHEI scores were inversely associated with abdominal obesity, hypertriglyceridemia, low high-density lipoprotein cholesterol, hyperglycemia, hypertension, and metabolic syndrome. Conclusion: Based on these results, we conclude that higher adherence to the KHEI was associated with lower metabolic syndrome risk factors and incidence of the metabolic syndrome.
Purpose: Extracellular signal-regulated kinase (ERK), which is part of the mitogen-activated protin kinase cascade, opposes initiation of the apoptotic cell death which is programmed by diverse cytotoxic stimuli. In this regard, the inhibition of ERK may be useful in improving the therapeutic efficacy of established anticancer agents. Materials and Methods: Murine hepatocarcinoma, HCa-I is known to be highly radioresistant with a TCD50 (radiation dose yield in $50\%$ cure) of more than 80 Gy. Various anticancer drugs have been found to enhance the radioresponse of this particular tumor but none were successful. The objective of this study was to explore whether the selective inhibition of MEK could potentiate the antitumor efficacy of radiation in vivo, particularly in the case on radioresistant tumor. C3H/HeJ mice hearing $7.5\~8\;mm$ HCa-I, were treated with PD98059(intratumoral injection of $0.16\;\mug/50\;\mul$). Results: Downregulation on ERK by PD98059 was most prominent 1h after the treatment. In the tumor growth delay assay, the drug was found to Increase the effect of the tumor radioresponse with an enhancement factor (EF) of 1.6 and 1.87. Combined treatment of 25 Gy radiation with PD98059 significantly increased radiation induced apoptosis. The peak apoptotic index (number on apoptotic nuclei in 1000 nuclei X100) was $1.2\%$ in the case of radiation treatment alone, $0.9\%$ in the case of drug treatment alone and $4.9\%,\;5.3\%$ in the combination treatment group. An analysis of apoptosis regulating molecules with Western blotting showed upregulation of p53, p$p21^{WAF1/CIP1}\;and\;Bcl-X_s$ in the combination treatment group as compared to their levels in either the radiation alone or drug alone treatment groups. The level of other molecules such as $Bcl-X_L4, Bax and Bcl-2 were changed to a lesser extent. Conclusion: The selective inhibition of MEK in combination with radiation therapy may have potential benefit in cancer treatment.
Journal of the Korea Organic Resources Recycling Association
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v.7
no.2
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pp.65-71
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1999
This study was conducted to evaluate the feasibility of food waste as a feed resources by fermentation and fermented food waste as a substitute of rat feed on the performance with measuring the liveweight gain, feed consumption, feed conversion and digestibility Sixty-two grams Sprague-Dawley line 36 rats were allocated three treatments 12 rats of each(3replication ${\times}$ 4 rats). The substitution level of fermented food waste to commercial broiler feed were control, 0: 100: treatment I, 10: 90: treatment II, 20:80. The chemical composition of fermented food waste was appeared to follows : dry matter, 88.47% : crude ash. 12.95: crude protein, 20.82%; crude fiber, 13.62; ether extract, 9.15%. The body weight of treatment I and II at 1 weeks was significantly lower than those of control(p<.05) and weekly weight gain of control at 0-1 weeks was significantly higher than those of treatment I and II(p<.05). Those were higher in treatment I than those of rest groups at 1-2 weeks(p<.05). Total weight gain of treatment II was significantly lower than those of control and treatment I(p<.05) Total feed consumption of treatment II was significantly higher than those of control (p<.05) and weekly feed consumption of control and treatment II at 3-4 weeks was significantly higher than those of treatment II(p<.05). but those were higher in treatment I and II than those of control at 2-3 weeks(p<.05). Commutative feed conversion of treatment II was significantly higher than those of control(p<.05) and weekly feed conversion of treatment II and III at 0-1 weeks was significantly higher than those of control(p<.05) Dry matter digestibility of control and treatment I was significantly higher than those of treatment II(p<.05) and organic matter digestibility was higher in control than those of treatment II(p<.05).
Park Kun-Koo;Jin Jung Sun;Park Ki Yong;Lee Yun Hee;Kim Sang Yoon;Noh Young Ju;Ahn Seung Do;Kim Jong Hoon;Choi Eun Kyung;Chang Hyesook
Radiation Oncology Journal
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v.19
no.2
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pp.171-180
/
2001
Purpose : Expression of TIMP, intrinsic inhibitor of MMP, is regulated by signal transduction in response to genotoxins and is likely to be an important step in metastasis, angiogenesis and wound healing after ionizing radiation. Therefore, we studied radiation mediated TIMP expression and its mechanism in head and neck cancer cell lines. Materials and Methods : Human head and neck cancer cell lines established at Asan Medical Center were used and radiosensitivity $(D_0)$, radiation cytotoxicity and metastatic potential were measured by clonogenic assay, n assay and invasion assay, respectively. The conditioned medium was prepared at 24 hours and 48 hours after 2 Gy and 10 Gy irradiation and expression of TIMP protein was measured by Elisa assay with specific antibodies against human TIMP. hTIMP1 promoter region was cloned and TIMP1 luciferase reporter vector was constructed. The reporter vector was transfected to AMC-HN-1 and -HN-9 cells with or without expression vector Ras, then the cells were exposed to radiation or PMA, PKC activator. EMSA was peformed with oligonucleotide (-59/-53 element and SP1) of TIMP1 promoter. Results : $D_0$ of HN-1, -2, -3, -5 and -9 cell lines were 1.55 Gy, 1.8 Gy, 1.5 Gt, 1.55 Gy and 2.45 Gy respectively. n assay confirmed cell viability, over $94\%$ at 24hrs, 48hrs after 2 Gy irradiation and over 73% after 10 Gy irradiation. Elisa assay confirmed that cells secreted TIMP1, 2 proteins continuously. After 2 Gy irradiation, TIMP2 secretion was decreased at 24hrs in HN-1 and HN-9 cell lines but after 10 Gy irradiation, it was increased in all cell lines. At 48hrs after irradiation, it was increased in HN-1 but decreased in HN-9 cells. But the change in TIMP secretion by RT was mild. The transcription of TIMP1 gene in HN-1 was induced by PMA but in HN-9 cell lines, it was suppressed. Wild type Ras induced the TIMP-1 transcription by 20 fold and 4 fold in HN-1 and HN-9 respectively. The binding activity to -59/-53, AP1 motif was increased by RT, but not to SP1 motif in both cell lines. Conclusions : We observed the difference of expression and activity of TIMPs between radiosensitive and radioresistant cell line and the different signal transduction pathway between in these cell lines may contribute the different radiosensitivity. Further research to investigate the radiation response and its signal pathway of TIMPs is needed.
Kim, Geum-Soog;Kim, Jung-Kon;Seong, Jae-Duck;Park, Chang-Kie;Suh, Hyung-Soo;Kwack, Yong-Ho
Korean Journal of Medicinal Crop Science
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v.4
no.2
/
pp.139-144
/
1996
This research was carried out to clarify effects of cultural condition on the content of Alisol B-monoacetate, whose function is antihypercolesterol in blood, and yield in Alisma Rhizomes. When the corm part of Alisma Rhizomes was extracted by shaking extraction at $25^{\circ}C$ and reflux extraction at $40^{\circ}C$ to $100^{\circ}C$ four times, the total content of Alisol B-monoacetate was 0. 402%, 0. 425% and 0. 402% at $25^{\circ}C$, $40^{\circ}C$ and $60^{\circ}C$ respectively. The recovery rate was 97% by three times extraction at $25^{\circ}C$, and 96% and 97% by three times extraction at $40^{\circ}C$ and $60^{\circ}C$ respectively. When the corm was harvested on Oct. 30, the content of Alisol B-monoacetate was 0. 46% but it was increased to 0. 71% on Nov. 30. In the case of Oct. 30, the corms of $S(4{\sim}14g\; FW)$ size were determined to contain the highest level of Alisol B-monoacetate (0. 53%), and the content was decreased as the corn size was increased. This tendency was also adopted on Nov. 30. On the other hand, the content of crude protein and starch was changed rarely by the period of harvest. When the planting depth was $0{\sim}1cm$, the yield was the highest as 206kg/10a, and the yield was decreased as the planting depth was deep. The variation of the content of Alisol B-monoacetate was small as 0. 33% to 0. 39% by planting depth. From the above results it could be concluded that the optimum time for harvest of Alisma Rhizomes is Nov. 30 and the optimum planting depth is $0{\sim}1cm$.
This experiment was conducted to assess genetic diversity of waxy corn inbred lines and to identify SSR markers related to major characteristics affected kernel quality for improving waxy corn $F_1$ hybrid with good quality. Diversity of 64 waxy com inbred lines was evaluated using 30 microsatellite markers. The 30 microsatellite markers representing 30 loci in the maize genome detected polymorphisms among the 64 inbred lines and revealed 225 alleles with a mean of 7.5 alleles per primer. The polymorphism Information content (PIC) value ranged from 0.14 to 0.87, with an average of 0.69. Based on Nei's genetic distances, the 64 inbred lines were classified into 9 groups by the cluster analysis. The group I included 26 inbred lines (41%), other groups included 3 to 9 inbred lines. One-way analysis of variance was conducted to identify significant relationship between individual markers and major characteristics that affect kernel quality. The analysis showed that umc1019 was related to amylopectin and crude protein content, me 1020 to amylopectin content and peak viscosity, and bnlg1537 to 100-kernel weight, kernel length, and kernel width.
This study aimed to investigate the anti-inflammatory effect of the ethanol extract from Chondrus ocellatus Holmes (COHEE) in RAW 264.7 cells and in a mouse ear edema model, by measuring the production of lipopolysaccharide-induced inflammatory response mediators. There were no cytotoxic effects on the proliferation of macrophages treated with COHEE compared with the control. COHEE inhibited the production of nitric oxide and pro-inflammatory cytokines [interleukin (IL)-6, tumor necrosis factor-α, and IL-1β]. The extract also reduced the expression of inducible nitric oxide synthase, cyclooxygenase-2, nuclear factor-κB p65, and phosphorylated mitogen-activated protein kinase in a dose-dependent manner. In the croton-oil-induced ear edema model, COHEE decreased the formation of mouse ear edema at the highest dose compared with the control, and histological analysis revealed that the epidermal/dermal tissue thickness and mast cell numbers were reduced. Therefore, these results suggest that COHEE may be a promising topical anti-inflammatory therapeutic material through its action of modulating NF-κB and the MAPK signaling pathway.
The anti-inflammatory effect of Sargassum patens C. Agardh ethanol extract (SPEE) was examined based on the lipopolysaccharide (LPS)-induced inflammatory response in this study. SPEE treatment was not cytotoxic to macrophages compared to the control. The production of NO was suppressed by SPEE by approximately 28% at $100{\mu}g/ml$, and levels of interleukin-6, tumor necrosis $factor-{\alpha}$, and $interleukin-1{\beta}$ decreased in a dose-dependent manner. In addition, the expression of inducible nitric oxide synthase, cyclooxygenase-2, and nuclear $factor-{\kappa}B$ was suppressed by SPEE treatment. In vivo, croton oil-induced mouse ear edema was attenuated by SPEE and the infiltration of mast cells into the tissue decreased. Based on these results, SPEE inhibits the release of LPS-induced pro-inflammatory cytokines and mediators, suggesting that SPEE is a potential agent for anti-inflammatory therapies.
Purpose : Human metapneumovirus(hMPV) is a respiratory viral pathogen that causes a wide spectrum of illnesses, ranging from asymptomatic infection to severe bronchiolitis. The virus has been identified world widely, but so far it has not been published in Korea. Methods : We obtained clinical samples by nasopharyngeal aspiration from 218 children hospitalized due to acute lower respiratory tract infections at Soonchunhyang University Hospital in Cheonan from October, 2004 to April, 2005. We designed specific primers from conserved region of fusion glycoprotein of hMPV. Total RNA was extracted and RT-PCR was performed, and single specific 423 bp product was obtained. The PCR product was confirmed to be fusion glycoprotein RNA by sequencing. Results : We detected hMPV in 15(6.9 percent) of the 218 hospitalized children. The infected children comprised nine boys and six girls; their mean age was 2.8 years(5 mo-12 yrs) and they were diagnosed with pneumonia(60 percent), bronchiolitis(33.3 percent), croup(6.6 percent). The number of cases of detected hMPV in Korea increased dramatically during the period from March to May 2005. Conclusion : hMPV is circulating in Korean children and is associated with respiratory tract infection. Additional studies are required to define the epidemiology and the extent of diseases in the general population caused by hMPV.
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