The high-level expression of a human single chain urokinase-type plasminogen activator (scu-PA) was achieved by employing a methotrexate (MTX)-dependent gene amplification system in Chinese hamster ovary (CHO) cells. By cotransfecting and coamplifying a scu-PA expression plasmid and dihydrofolate reductase (DHFR) minigene, several scu-PA expressing CHO cell lines were selected and gene-amplified. These recombinant cell lines, NGpUKs, secreted a completely processed scu-PA of 54 kD and up to 60mg/L was accumulated in the culture medium when they were adapted to an optimal MTX concentration. Over 95% of the scu-PA expressed was secreted in the culture medium and identified having the proper function of a plasminogen activator when activated by plasmin. Based on a genomic Southern analysis, a representative subclone, MGpUK-5, exhibited MTX-dependent scu-PA gene amplification, plus the initial single-copy gene of scu-PA eventually turned into about 150 copies of the amplified gene of scu-PA after gradual adaptation to 2.0$\mu$M of MTX. Meanwhile, the transcripts kof the scu-PA gene increased, although -early saturation of transcription was identified at 0.1$\mu$M of MTX. The scu-PA production by the MGpUK-5 subclone also increased relative to the gene amplification and increased transcripts, however, the relationship was not linearly proportional. Accordingly, since the MGpUK cell lines expressed elevated levels of enzymatically active scu-PA, these cell lines could be applied to the largescale production of scu-PA.
The Journal of the Convergence on Culture Technology
/
v.6
no.3
/
pp.159-165
/
2020
This study was attempted to identify the status of the work of the PA and provide the direction and evidence of policy making on the PA scheme. A convergence study was conducted to analyze a total of 23 articles of papers and Internet news through systematic review methodology. A literature search was performed using KISS, NDSL, RISS, National assembly library, and 11 major domestic daily newspapers and Youth Medical Newspaper. The analysis of eight papers showed that the PA's work-related exhaustion, job stress, unclear scope of work and lack of regulations reduced job satisfaction. Through 15 press articles, it was understood that it was urgent to establish a legal system and education system for qualifications and work as professional medical personnel amid a conflict of positive and negative opinions of related agencies. In order to increase the utilization of PA in the future, it is necessary to secure institutional devices suitable for Korean affairs and prepare educational programs.
Objective : This study has been carried out to understand the effect of several herbal medicine concentrated solution on the hyperglycemic mice Induced with streptozotocin(STZ). Methods : The 60 mg/kg of STZ injection into mice twice by 24 h interval and then 120 mg/kg of STZ injection again 3 days after the earlier injection. Control group was subjected to natural recovery, however, treated groups were fed 0.2 ml of several herbal medicine-concentrated solution (PA (x2, several herbal medicine-concentrated solution 1 group); PB (x4 several herbal medicine-concentrated solution 2 group) daily for 6 weeks. Result : The weight of PA was higher than that of control, but weight of PB was lower than control. The blood level of control increased continuously, reaching to 350mg/dL after 6 weeks, however, PA and PB showed a fast reduction of blood glucose. In blood glucose tolerance test, PA and PB showed better resistance than control. The GOT level in significantly(p<0.05) decreased in PA and PB compared with control group. The BUN and creatinine levels are significantly(p<0.01) decreased in PA compared with control group. Feeding of several herbal medicine-concentrated solution in a concentration of PA had an efficient effect on regeneration or recovery of Langerhans islet and ${\beta}-cell$ damaged by STZ. More Langerhans islet and high insulin-immunohistochemical resistance were observed in PA compared with control, but they were higher in PB than in PA. The number of Langerhans islet ${\beta}-cell$ and Langerhans islet. Conclusions : The result from the six weeks of observation demonstrates that the several herbal medicine concentrated solution have a positive effect of lowering the level of blood sugar and they increased insulin concentration. They have an effect for recovery of pancreas tissue and recovery of kidney, liver function from a diabetes mellitus.
Phytic acid(PA) (Inositol hexaphosphate, $IP_6$) is a naturally occurring polyphosphorylated carbohydrate that is present in substantial amounts in almost all plants and mammalian cells. Recently PA has received much attention for its role in anticancer activity. In the present study, the preventive effects of PA on colon carcinogenesis were investigated. Six-week old Fisher 344 male rats were fed a AIN-93G purified diet and PA(0.5% or 2% PA in water) for 8 weeks. The animals received two ($1^{st}\;and\;2^{nd}$ week) injections of azoxymethane(AOM, 15 mg/kg b.w.) to induce colonic aberrant crypt foci(ACF). After sacrifice, the total numbers of aberrant crypts(AC) and ACF in colonic mucosa were examined after staining with methylene blue. Blood and serum were analyzed with a blood cell differential counter and an automatic serum analyzer. AOM induced the total numbers of $142.3{\pm}22.3$ ACF/colon and $336.6{\pm}55.1$ AC/colon. PA at the doses of 0.5 and 2% decreased the numbers of ACF and AC/colon in a dose-dependent manner. The numbers of ACF/colon and AC/colon by PA at the dose of 0.5% were $124.4{\pm}28.5\;and\;302.7{\pm}67.3$, respectively. PA at the dose of 2% significantly decreased the ACF and AC numbers to $109{\pm}18.1\;and\;254.8{\pm}50.6$, respectively(p<0.01). Especially, 2% PA significantly reduced the number of large ACF(${\geq}4$ AC/ACF) from $26.8{\pm}6.2$ ACF/colon to $15{\pm}6.7$ ACF/colon(p<0.01). Although some parameters in blood counts and serum chemistry were changed compared with the control, no specific toxicity was found. These findings suggest that phytic acid can be a chemopreventive agent for colon carcinogenesis resulting from inhibition of the development of ACF in the F344 rat.
The present study was conducted to evaluate fertigation effects of balanced manure nutrients from combined fertilizers on the growth of eggplant (Solanum melongena L.) and soil chemical properties in greenhouse cultivation. There are 4 treatments for fertigation; animal liquid manure (LM), LM supplemented with chemical ferilizer (LM+CF) to make nutrient-balanced ferilizer, LM supplemented with chemical ferilizer and phosphorus acid (LM+CF+PA), and conventional chemical fertilzer (CCF). Fruit lenth, width, and weight for LM treatment were significantly lower than LM+CF, LM+CF+PA, and CCF treatments. There was no significant differences in fruit length, width, and weight among LM+CF, LM+CF+PA, and CCF treatments. The yield of eggplants per plant for LM treatment (26.2 kg) was decreased by 6% compared to CCF treatment (27.9 kg). The growth and yield of eggplants were not significant different among CCF, LM+CF, and LM+CF+PA treatments. The marketable yield for LM treatment was lowest among 4 treatments and that for LM+CF+PA treatment (26.6 kg) was increased by 8% compared to CCF treatment (24.1 kg). The chemical properties of the soil for CCF treatment, in general, tended to be higher compared to the other treatments. In conclusion, the present study demonstrates that the application of balanced liquid manure combined with mineral fertilizer to soil is considered as a good management practices because it improves, eggplant quality and soil properties.
Jung, Seung Hun;Kang, Ji Hoon;Park, Seung Jong;Seong, Ki Hyun;Song, Kyung Bin
Journal of the Korean Society of Food Science and Nutrition
/
v.43
no.6
/
pp.901-908
/
2014
Blueberries and strawberries are highly perishable and easily contaminated with microorganisms. To maintain the quality of these commodities during refrigerated storage, the effects of two packaging materials as well as passive modified atmosphere packaging on the quality of blueberries and strawberries were investigated. The harvested blueberries and strawberries were first treated with combined non-thermal treatment of aqueous chlorine dioxide and fumaric acid, followed by packaging with polyolefin film (6,000 mL $O_2/m^2{\cdot}24hr{\cdot}atm$ at $24^{\circ}C$) and polyamide/polyamide/polyethylene film (PA/PA/PE, 60 mL $O_2/m^2{\cdot}24hr{\cdot}atm$ at $24^{\circ}C$), respectively. After combined sanitizer treatment, the populations of total aerobic bacteria in blueberries and strawberries were reduced by 2.50 and 1.97 log CFU/g while those of yeast and molds were reduced by 1.95 and 2.18 log CFU/g, respectively, compared with the control. In particular, microbial growth in these samples packed with PA/PA/PE film was reduced during refrigerated storage. In addition, the blueberries and strawberries packed with PA/PA/PE film underwent lower weight loss than those packed with polyolefin film during storage. These results suggest that appropriate packaging with proper gas permeability is necessary to maintain the quality of blueberries and strawberries during refrigerated storage.
Park, Sora;Thomas, Shalom Sara;Cha, Youn-Soo;Kim, Kyung-Ah
Journal of Nutrition and Health
/
v.53
no.6
/
pp.583-595
/
2020
Purpose: This study examined the effects of Sargassum horneri extracts on palmitic acid (PA)-induced endoplasmic reticulum (ER) stress in HepG2 cells. Methods: HepG2 cells were treated with varying concentrations of S. horneri extract or PA, and the cell viability was measured by water soluble tetrazolium salts analysis. The effective induction of ER stress and the effects of S. horneri were investigated through an examination of the ER stress-related genes, such as activating transcription factor 4 (ATF4), X-box binding protein (XBP1s), C/EBP homologous protein (CHOP), and 78-kDa glucose-regulated protein (GRP78) by quantitative reverse transcription polymerase chain reaction. The expression and activation levels of unfolded protein response (UPR) associated proteins, such as inositol-requiring enzyme-1α (IRE1α), eukaryotic translation initiation factor 2 alpha submit (eIF2α), and CHOP were examined by western blot analysis. Results: The treatment with PA increased the expression of UPR associated genes significantly and induced ER stress in a 12-hour treatment. Subsequent treatment with S. horneri reduced mRNA expression of ATF4, GRP78, and XBP1s. In addition, the protein levels of phosphate (p)-IRE1α, p-elF2α, and CHOP were also reduced by a treatment with S. horneri. An analysis of sirtuin (SIRT) mRNA expression in the S. horneri and PA-treated HepG2 cells showed that S. horneri increased the levels of SIRT2, SIRT6, and SIRT7, which indicates a possible role in reducing the expression of ER stress-related genes. Conclusion: These data indicate that S. horneri can exert an inhibitory effect on ER stress caused by PA and highlight its potential as an agent for managing various ER stress-related diseases.
Anthrax caused by Bacillus anthracis is one of the most important zoonotic diseases. The bacterium produces several virulence factors. Of the factors, protective antigen (PA) of tripatite toxin has been identified as a central component in the pathogenesis of anthrax. However, precise roles of PA and other cellular components in the reaction with the target cells remain to be elucidated, especially in the initial stage of the disease. Three B anthracis antigens were prepared for investigation; PA, sonicated cellular antigens (S-Ag) and formalin-inactivaed whole cell antigens (W-Ag). PA was purified from culture supernatant of the bacterium using FPLC system with MonoQ. S-Ag and W-Ag were prepared by sonication and formalin inactivation of the cultured cells, respectively. Purity of the antigens was confirmed by SDS-PAGE and Western blot analysis. The roles of these antigens in the production of inflammatory mediators such as NO, IL-6 and $TNF{\alpha}$ from mouse peritoneal macrophages were investigated. PA alone did not induce the production of the inflammatory mediators while the other antigens, S-Ag and W-Ag, did in a dose and time dependent manner. These results suggested that in addition to major virulence factors, other cellular antigens are also involved in the initial stage of the disease by the induction of inflammatory mediators.
This study aims at investigating the effect of soil moisture control and truss limited high density culture on the growth, fruit yield and quality of tomato. To minimize of loss yield of tomato, flower cluster in number was limited to two and three truss and planting density was raised. Soil moisture control was started from 40 days after anthesis and irrigation point was set in -30kPa and -50kPa, which were compared with -10kPa For high density culture, the planting number of truss limited high density culture was planted twice as many as control. Soil moisture repression reduced the growth of stem diameter, leaf and plant height. Leaf chlorophyll content was higher in -50kPa and -30kPa than control. No significant differences, however, shows in -10kPa. The occurrence rate of bloom-end rot and cracking was increased by growing of irrigation repression. Pinching three fruit truss was higher than pinching two fruit truss in the occurrence rate of them. Soil moisture repression resulted in the reduction of fruit weight and in special, truss limited high density was distinctly decreased in -50kPa. The number of fruit was not affected by soil moisture control, but 3rd flower cluster was lower than 2nd flower cluster in the number of fruits and 2nd one was lower than 1st one. Under irrigation repression, rate of dry matter tended to grow in -30kPa, -50kPa compared with control and pinching three fruit truss was higher than two truss. Marketable yield dropped to 36.7%m 46.3 in -30kPa, -50kPa on pinching two fruit truss and dropped to 27.3%, 32.3% in 3rd flower cluster compared with control.
This study was conducted to investigate effects of irrigation points on cucumber growth and water saving under greenhouse. Automatic irrigation system and tensiometers were installed at four plots to measure soil water tension and properties of irrigated water. Each plot was irrigated at different irrigation points; soil water tension of 15, 20, 30, and 40 kPa, respectively. Conventional irrigation plot without tensiometer was also investigated. The total yield and sugar contents of cucumber had no significant differences between plots. However, irrigated water volumes were saved most when irrigation points were 30, and 40 kPa. The two plots of 30, and 40 kPa treatments used about 60% less of irrigated water than the plot of conventional practice did. In conclusion, this research suggested that the optimum irrigation point for cucumber cultivation in greenhouse can be soil water tension of 30 kPa based on the results of overall cucumber quality, and Greenhouse water usage.
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