Choi, Yu Jin;Park, Su Hyun;Kim, Ji Su;Wi, Soo Jin;Park, Ky Young
Journal of Plant Biotechnology
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v.40
no.3
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pp.169-177
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2013
Recently, the number of people with diabetes is rapidly increasing, coupled with the fact that the insulin market is remarkably increasing. Therefore, molecular farming for plant-derived pharmaceutical protein production is reported as becoming more attractive than ever. In this study, we carried out experiments step by step for development of recombinant insulin constructs, which were transformed into E. coli system, in vitro transcription and translation system, and tobacco cells. At first, recombinant proinsulin protein was successfully produced in in vitro transcription and translation system with wheat germ extract. After which, recombinant construct of prominiinsulin encoded a fusion protein of 7.8 kDa with trypsin cleavage sites at N terminus and C terminus of minimized C-peptide was tried to in vitro expression using E.coli culture. After purification with His-tag column, the resulting recombinant prominiinsulin protein was processed with trypsin, and then checked insulin biosynthesis by SDS-PAGE and western blot analysis with anti-insulin monoclonal antibody. The immunoreactive product of trypsin-treated miniinsulin was identical to the predicted insulin hexamer. The construct of 35S promoter-driven preprominiinsulin recombinant gene with signal peptide region for ER-targeting and red fluorescence protein gene [N terminus ${\rightarrow}$ tobacco E2 signal peptide ${\rightarrow}$ B-peptide (1-29 AA) ${\rightarrow}$ AAK ${\rightarrow}$ A-peptide (1-21 AA) ${\rightarrow}$ RR ${\rightarrow}$ His6 ${\rightarrow}$ KDEL ${\rightarrow}$ C terminus] was transformed into BY-2 tobacco cells. A polypeptide corresponding to the 38-kDa molecular mass predicted for fusion protein was detected in total protein profiles from transgenic BY-2 cells by western analysis. Therefore, this recombinant preprominiinsulin construct can be used for generation of transgenic tobacco plants producing therapeutic recombinant insulin.
Lee, Wi Young;Na, Sung June;Park, Eung-Jun;Han, Sang Urk
Korean Journal of Plant Resources
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v.27
no.4
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pp.279-285
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2014
Nutritional composition, including total phenolics, tannins and nutrient components, of acorns of Q. actissima, Q. serrata, Q. variabilis and Q. mongolica were analyzed. Acorns were collected from each tree species, which were grown in a seed orchard. Contents of both total phenolics and tannins in acorns of Q. serrata were higher than those of Q. actissima (p<0.05). Interestingly, Q. serrata contained the highest amount of water soluble tannins (71 mg/g dw) and the lowest levels of water insoluble tannins (8.1 mg/g dw) among 4 oak species, resulting that acorns of Q. serrata had the lowest proportion of insoluble tannins. Among 4 oak species tested, Q. mongolicav acorns contained the highest levels of both total dietary fiber (TDF) and ascorbic acid, while the content of beta-carotene in Q. mongolicav was 52-fold lower than that in Q. variabilis ($520{\mu}g/100g$). Our result showed that nutritional composition of acoms was significantly different between oak species, indicating that tastes or nutritional values might be different as well among major oak species in Korea.
Background: Congenital cystic diseases of the lung are uncommon, and they share similar embryogenic and clinical characteristics. But they are sometimes vary widely in their presentation and severity. Therefore they are often difficult to make different diagnosis each other, and all require surgical treatment. Material and Method: From 1993 to 2006, 38 patients underwent surgical procedures under these diagnostic categories in the Depart. of Thoracic and. Cardiovascular Surgery, Busan-Paik Hospital, College of Medicine, Inje University. And we retrospectively reviewed these patients' charts for clinical presentations, surgical procedures, pathologic findings and postoperative morbidity and mortality. Result: There were 22 males and 16 females, ages ranged from 1 month after birth to 51 years and mean age was 20.8 years. The main symptoms were 19 fever, cough, sputum production due to recurrent infection, 7 dyspnea, 8 chest discomfort, 4 hemoptysis, but eight patients were asymptomatic. Computed tomography was chosen as diagnostic modalities and available for operation plan for all of patients. For all the cases, surgical resection were performed. Lobectomy was performed in 28 patients, simple excision (resection) in 8 patients, segmentectomy or wedge resection in 2 patients. There were 10 pulmonary sequestrations, 15 congenital cystic adenomatoid malformations (CCAM), 11 bronchogenic cysts, and 2 congenital lobar emphysemas. They all were confirmed by pathologic exams. The complications were 6 wound disruption or infection, 2 chylothorax, 1 ulnar neuropathy, but all of them were resolved uneventful. There was no persistent air leakage, respiratory failure, operative mortality and recurrence. Conclusion: We performed immediate surgical removal of congenital cystic lung lesions after diagnosis and obtained good results, so reported them with literature review.
Journal of the Korean Society of Food Science and Nutrition
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v.41
no.8
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pp.1079-1085
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2012
Obesity is an important issue worldwide as it may associated with increased prevalence of metabolic diseases. Mung bean is known as a functional food for decreasing the glycemic index and lipid profile of plasma. The purpose of this study was to investigate the anti-obesity effects of vitexin from mung bean on the regulation of adipocyte differentiation and adipocytokine secretion. When 3T3-L1 adipocytes were treated with vitexin from days 0 to 14 at various levels of 25, 50, 100, and $200{\mu}M$, there was no change in cell viability. Vitexin treatment at 50, 100, and $200{\mu}M$ decreased triacylglycerol levels in cells, but only $100{\mu}M$ vitexin induced lipolysis. At $200{\mu}M$ of vitexin, phosphorylation of p38 and ERK, which causes secretion of inflammatory adipocytokines, was depressed, whereas there was an increase in expression of $PPAR{\gamma}$, the key regulator of adipocyte differentiation. Phosphorylation of AMPK increased at $100{\mu}M$ vitexin. TNF-${\alpha}$ and aP2 mRNA expression increased at $25{\mu}M$ vitexin, whereas only TNF-${\alpha}$ mRNA expression increased at $200{\mu}M$ vitexin. Further, the mRNA levels of TNF-${\alpha}$ and aP2 decreased at other concentrations in a dose-dependent manner. Since we observed that mRNA expression of C/EBP, SREBP1, and $PPAR{\gamma}$ did not change upon vitexin treatment, our future studies will investigate other genes such as mTOR, which is related with apoptosis signaling, or SIRT1, which is associated with inhibition of adipogenesis. Our results indicate that vitexin at concentrations between 100 and $200{\mu}M$ is suitable in vivo for the development of mung bean as an anti-obesity therapy or functional food.
Journal of the Korean Society of Food Science and Nutrition
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v.42
no.9
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pp.1499-1512
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2013
The purpose of this study was to compare the coffee shop visits of Koreans and Mongolians, and to determine the quality attributes that should be managed by Importance-Performance Analysis (IPA). The survey was conducted in Seoul and the Gyeonggi Province of Korea, and at Ulaanbaatar in Mongolia from April to May 2012. The questionnaire was distributed to 380 Koreans and 380 Mongolians, with 253 and 250 responses from the Koreans and Mongolians, respectively, used for statistical analyses. From the results, Koreans visited coffee shops more frequently than Mongolians, with both groups mainly visiting a coffee shop with friends. Koreans also spent more time in a coffee shop than Mongolians. In addition, they generally used a coffee shop, regardless of time. In terms of coffee preference, Koreans preferred Americano and Mongolians preferred Espresso. The most frequently stated purpose of Koreans for visiting a coffee shop was to rest, while Mongolians typically visited to drink coffee. The general price range respondents spent on coffee was less than 4~8 thousand won for the Koreans and 2~4 thousand won for the Mongolians. Both Koreans and Mongolians obtained information about coffee shops from recommendations. According to the IPA results of 20 quality attributes of coffee shops, the selection attributes with high importance but low satisfaction were quality, price, and kindness for Koreans, but none of the attributes was found for Mongolians.
Purpose: The menstrual cycle of normal premenopausal patients was divide into menstrual flow phase, proliferative phase, ovulatory phase, secretory phase. The aim of this study was to ovarian and endometrial $^{18}F$-FDG uptake during the menstrual cycle in normal premenopausal patients. Materials and Methods: We identified 200 incidental $^{18}F$-FDG uptake in the ovary. The patient fasted at least 6 hours before receiving an intravenous injection of 370-592 MBq (10-16mCi) of $^{18}F$-FDG. Scanning from the base of the skull though the mid thigh was performed using the Discovery Ste PET/CT system (GE Healthcare, Milwaukee, WI, USA). Ovarian and endometrial $^{18}F$-FDG uptake (expressed as standardized uptake value) was measured on PET/CT image. Results: Two peaks of increased endometrial $^{18}F$-FDG uptake were identified during the menstrual cycle. The $SUV_{avg}$ and $SUV_{max}$ was $2.89{\pm}1.04$ and $3.17{\pm}1.59$ in menstruating patients, $2.4{\pm}0.88$ and $2.98{\pm}1.14$ in proliferative phase patients, $3.59{\pm}1.76$ and $3.17{\pm}1.67$ in ovulatory phase patients, $2.58{\pm}1.39$ and $3.1{\pm}1.8$ in secretory phase patients. Conclusions: Increased ovarian and endometrial $^{18}F$-FDG uptake could be found the time of menstrual flow and ovulatory phase of menstrual cycle. Increased uptake in endometrial adjacent to a cervical tumor does not necessarily reflect endometrial tumor invasion. Since increased uptake was dependent on the menstrual cycle, it can be avoided by scheduling PET/CT just after menstruation. Non-menstrual-related endometrial uptake may be instrumental in establishing a diagnosis in a premenopaual patient.
Park, Sang Sun;Kim, Eun Joung;Son, Chang Young;Wi, Jeong Ook;Park, Kyung Hwa;Cho, Gye Jung;Ju, Jin Young;Kim, Kyu Sik;Kim, Yu II;Lim, Sung Chul;Kim, Young Chul;Park, Kyung Ok;Na, Kook Joo
Tuberculosis and Respiratory Diseases
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v.55
no.1
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pp.88-97
/
2003
Background : Although smoking is a major cause of chronic obstructive pulmonary disease (COPD), only 10-20% of cigarette smokers develop symptomatic COPD, which suggests the presence of genetic susceptibility. This genetic susceptibility to COPD might depend on variations in the activities of the enzyme that detoxify hazardous chemical products, such as microsomal epoxide hydrolase (mEPHX) and glutathione-S transferase M1 subunit (GSTM1) genes. Methods : The genotypes of 58 patients with COPD, and 79 age matched control subjects, were determined by a polymerase chain reaction, followed by restriction fragment length polymorphism (PCR-RFLP) for the mEPHX, and multiplex PCR for the GSTM1. Results : GSTM1 was deleted in 53.3% of the subjects. There was no difference in GSTM1 deletion rates between the COPD patients (32/58, 55.2%) and the control subjects (41/79, 51.9%). The combination patterns of two polymorphisms of mEPHX showed slow enzyme activity in 29(21.2%), normal in 73(53.3%) and fast in 32(23.4%). The COPD group (7/57, 12.3%) showed a significantly lower incidence of slow enzyme activity compared to the control subjects (22/77, 28.6%, p<0.05). However, when the COPD and control groups were compared with smokers only, there were no significant differences in the genotypes of GSTM1 and mEPHX. Conclusion : The genotypes of GSTM1 and mEPHX were not significant risk factors of COPD in this cohort of study.
Purpose: Much evidence suggests long-term cigarette smoking alters coronary vascular endothelial response. On this study, we applied nonnegative matrix factorization (NMF), an unsupervised learning algorithm, to CO-less $H_2^{15}O-PET$ to investigate coronary endothelial dysfunction caused by smoking noninvasively. Materials and methods: This study enrolled eighteen young male volunteers consisting of 9 smokers $(23.8{\pm}1.1\;yr;\;6.5{\pm}2.5$ pack-years) and 9 nonsmokers $(23.8{\pm}2.9 yr)$. They do not have any cardiovascular risk factor or disease history. Myocardial $H_2^{15}O-PET$ was performed at rest, during cold ($5^{\circ}C$) pressor stimulation and during adenosine infusion. Left ventricular blood pool and myocardium were segmented on dynamic PET data by NMF method. Myocardial blood flow (MBF) was calculated from input and tissue functions by a single compartmental model with correction of partial volume and spillover effects. Results: There were no significant difference in resting MBF between the two groups (Smokers: 1.43 0.41 ml/g/min and non-smokers: $1.37{\pm}0.41$ ml/g/min p=NS). during cold pressor stimulation, MBF in smokers was significantly lower than 4hat in non-smokers ($1.25{\pm}0.34$ ml/g/min vs $1.59{\pm}0.29$ ml/gmin; p=0.019). The difference in the ratio of cold pressor MBF to resting MBF between the two groups was also significant (p=0.024; $90{\pm}24%$ in smokers and $122{\pm}28%$ in non-smokers.). During adenosine infusion, however, hyperemic MBF did not differ significantly between smokers and non-smokers ($5.81{\pm}1.99$ ml/g/min vs $5.11{\pm}1.31$ ml/g/min ; p=NS). Conclusion: in smokers, MBF during cold pressor stimulation was significantly lower compared wi4h nonsmokers, reflecting smoking-Induced endothelial dysfunction. However, there was no significant difference in MBF during adenosine-induced hyperemia between the two groups.
We investigated the effects of rising temperatures on the photosynthesis, mineral composition, and growth of radish (Raphanus sativus var. hortensis) in a winter cropping system using a temperature gradient tunnel to predict the impact of rising global temperatures. Vegetative growth, including shoot and root fresh and dry weights, shoot length, and root length and diameter, was high under elevated temperatures (ambient $+4^{\circ}C$ and $+7^{\circ}C$) compared with ambient temperature. At elevated temperatures, the N, P, Ca, Mg, and Fe contents were high in shoots, whereas in roots, the K, Ca, Mg, and Fe contents were high and the Cu content was low. The maximum photosynthetic rates ($22.1{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ at ambient temperature $+4^{\circ}C$ and $22.9{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ at ambient temperature $+7^{\circ}C$) at elevated temperatures were more than twice that ($9.7{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) at ambient temperature, whereas the water use efficiency was lower at elevated temperatures. These results suggest that rising global temperatures will lead to increased mineral absorption and photosynthesis in radish in winter cropping systems, subsequently favoring plant growth, although the water requirements will be high.
Kim, Jae-Young;Lee, Jin-Young;Lee, Wi-Young;Yi, Yong-Sub;Lim, Yoong-Ho
Microbiology and Biotechnology Letters
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v.38
no.4
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pp.414-419
/
2010
Plants extracts are good resources to find functional compounds for human health. The following eight plants were collected and total phenolic contents were determined. Acer psedo-siebolianum showed the highest phenolic contents, 16.4 mg/g, whereas Cercidiphyllum japonica showed the lowest contents, 1.9 mg/g. The DPPH free radical scavenging capacities of the plant extracts showed high activity in following order : Acer ginnala ($21.3\;{\mu}g/mL$) > Cornus walteri ($23.9\;{\mu}g/mL$) > Distylum racemosum ($29.2\;{\mu}g/mL$) > Castanopsis cuspidata var. Thunbergii ($31.7\;{\mu}g/mL$) > Acer psedo-siebolianum ($34.6\;{\mu}g/mL$) > Thuijopsis dolabrata cv. Aurea ($53.1\;{\mu}g/mL$) > Cercidiphyllum Japonica ($115.2\;{\mu}g/mL$). Also the mushroom tyrosinase inhibitory activities of total extracts were determined at different concentration. D. racemosum extract showed highest (49.1% at 1,000 mg) in inhibitory activity than other seven extracts. The ethanol fraction $IC_{50}$ value: $118.1\;{\mu}g/mL$) from D. racemosum showed more inhibitory activity than ethyl acetate fraction ($IC_{50}$ value: $203\;{\mu}g/mL$). The ethanol fraction on showed no significant cytotoxicity in B16/F1 cells line up to $60\;{\mu}g/mL$. Over $80\;{\mu}g/mL$ of ethanol fraction showed cytotoxicity in B16/F1 cells. The melanin contents of cells were significantly attenuated by ethanol fraction in a dose-dependent manner. The $IC_{50}$ value of ethanol fraction was $75.4\;{\mu}g/mL$.
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