For the production and purification of a single chain human insulin precursor, four types of fusion peptides $\beta$-galactosidase (LacZ), maltose binding protein (MBP), glutathione-S-transferase (GST), and (His)(sub)6-tagged sequence (HTS) were investigated. Recombinant E. coli harboring hybrid genes was cultivated at 37$\^{C}$ for 1h, and gene induction occurred when 0.2mM of isopropyl-D-thiogalactoside (IPTG) was added to the culture broth, except for E. coli BL21 (DE3) pLysS harboring a pET-BA cultivation with 1.0mM IPTG, followed by a longer than 4h batch fermentation respectively. DEAE-Sphacel and Sephadex G-200 gel filtration chromatography, amylose affinity chromatography, glutathione-sepharose 4B affinity chromatography, and a nickel chelating affinity chromatography system as a kind of immobilized metal ion affinity chromatography (IMAC) were all employed for the purification of a single chain human insulin precursor. The recovery yields of the HTS-fused, GST-fused, MBP-fused, and LacZ-fused single chain human insulin precursors resulted in 47%, 20%, 20%, and 18% as the total protein amounts respectively. These results show that a higher recovery yield of the finally purified recombinant peptides was achieved when affinity column chromatography was employed and when the fused peptide had a smaller molecular weight. In addition the pET expression system gave the highest productivity of a fused insulin precursor due to a two-step regulation of the gene expression, and the HTS-fused system provided the highest recovery of a fused insulin precursor based on a simple and specific separation using the IMAC technique.
This study was performed to produce succinic acid from biomass by developing mutants of Cellulomonas flavigena in which the succinate dehydrogenase gene (sdh) is deleted. For development of succinate producing mutants, the upstream and downstream regions of sdh gene from C. flavigena and antibiotic resistance gene (neo, bla) were inserted into pKC1139, and the recombinant plasmids were transformed into Escherichia coli ET12567/pUZ8002 which is a donor strain for conjugation. C. flavigena was conjugated with the transformed E. coli ET12567/pUZ8002 to induce the deletion of sdh in chromosome of this bacteria by double-crossover recombination. Two mutants (C. flavigena H-1 and H-2), in which sdh gene was deleted in the chromosome, were constructed and confirmed by PCR. To estimate the production of succinic acid by the two mutants when the culture broth was fermented with biomass such as CMC, xylan, locust gum, and rapeseed straw; the culture broth was analyzed by HPLC analysis. The succinic acid in the culture broth was not detected as a fermentation products of all biomass. One of the reasons for this may be the conversion of succinic acid to fumaric acid by sdh genes (Cfla_1014 - Cfla_1017 or Cfla_1916 - Cfla_1918) which remained in the chromosomal DNA of C. flavigena H-1 and H-2. The other reason could be the conversion of succinyl-CoA to other metabolites by enzymes related to the bypass pathway of TCA cycle.
This study aimed to determine the minimum period of aeration treatment of co-digestate to develop it as liquid fertilizer and the chemical changes that occur in the aerobic liquefying process. The co-digestates were divided into three types depending on their additives: swine slurry anaerobic digestate (SS AD), swine slurry 70% + cow slurry 30% anaerobic digestate (SS + CS AD), and swine slurry 70% + apple pomace 30% anaerobic digestate (SS + AP AD). The pH of all co-digestates increased rapidly after 3 days of aerobic treatment, but had slightly decreased in SS AD after 9 days and in SS + CS AD and SS + AP AD after 15 days. All co-digestates showed a strongly reduced pH between 27 and 36 days of aeration treatment. SS AD had lower pH value, dissolved oxygen (DO), NH4-N, and NO3-N content under aerobic conditions than other co-digestates. To assess the fully decomposed liquid fertilizer, a germination test was performed on the undiluted and diluted co-digestate using the liquid fertilizer germination index (LFGI) method. The relative germination ratio, relative root elongation, and germination index of SS AD were higher than those of the others. When the LFGI method was used for the germination test, all co-digestates showed an appropriate germination index of 70 after 60 days of aeration treatment. Thus, we suggest that the minimum period of aeration treatment for co-digestates might be 60 days to develop the fully decomposed liquid fertilizer.
Journal of the Institute of Convergence Signal Processing
/
v.12
no.3
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pp.157-162
/
2011
This study is about implementing wireless transferring system in pre-hospital cardiopulmonary resuscitation(CPR). Also, this study includes monitoring based feedback between patient and hospital to increase the survival rate of emergency patient by developing the performance of cardiopulmonary resuscitation in pre-hospital. It minimizes the loss of flow rate or gastric inflation through the space between the airway and the esophagus, which enables the inspiration-expiration rate to be measured more precisely. Due to these reasons this study applied ET insertion based respiratory sensor to measure flow rate. The main indices of artificial ventilation are justified from minute respiration(V), end-tidal $CO_2(E_TCO_2)$, and tracheal pressure($P_{tr}$). The simulation is performed to verify the bandwidth and delay time of transport network for in-hospital monitoring even as transporting images and voice information simultaneously. The total bandwidth is 815 kbps, and WLAN (IEEE 802.11x) is used as communication protocol. The network load is under 1.5% and the transmit delay time is measured under 0.3 seconds.
The Hayabusa 2 mission target asteroid (162173) Ryugu is a near-Earth, carbonaceous (C-type) asteroid. Before the arrival, this asteroid is expected to be covered with mm- to cm- sized grains through the thermal infrared observations [1]. These grains are widely understood to be formed by past impacts with other celestial bodies and fractures induced by thermal fatigue [2]. However, the close-up images by the MASCOT lander showed lumpy boulders but no abundant fine grains [3]. Morota et al. suggested that there would be submillimeter particles on the top of these boulders but not resolved by Hayabusa 2's onboard instruments [4]. Hence, we conducted polarimetry of Ryugu to investigate microscopic grain sizes on its surface. Polarimetry is a powerful tool to estimate physical properties such as albedo and grain size. Especially, it is known that the maximum polarization degree (Pmax) and the geometric albedo (pV) show an empirical relationship depending on surface grain sizes [5]. We observed Ryugu from UT 2020 November 30 to December 10 at large phase angles (ranging from 78.5 to 89.7 degrees) to derive Pmax. We modified TRIPOL (Triple Range Imager and POLarimeter, [6]) to attach to the 1.8-m telescope at the Bohyunsan Optical Astronomy Observatory (BOAO). With this instrument, we observed the asteroid and determined linear polarization degrees at the Rc-band filter. We obtained sufficient data sets from 7 nights at this observatory to determine the Pmax value, and collaborated with other observatories in Japan (i.e., Hokkaido University, Higashi-Hiroshima, and Nishi-Harima) to acquire linear polarization degrees of the asteroid from total 24 nights observations with large phase angle coverage (From 28 to 104 degrees). The observational results have been published in Kuroda et al. (2021) [7]. We thus found the dominance of submillimeter particles on the surface of Ryugu from the comparison with other meteorite samples from the campaign observation. In this presentation, we report our activity to modify the TRIPOL for the 1.8-m telescope and the polarimetric performance. We also examine the rotational variability of the polarization degree using the TRIPOL data.
Kim, Chi Nyon;Lee, Se Hoon;Kim, Hyun-Soo;Youn, Young-Shik;Roh, Jaehoon
Journal of Korean Society of Occupational and Environmental Hygiene
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v.11
no.2
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pp.118-125
/
2001
Recently, biochemical analysis using hemoglobin adduct is frequently performed to evaluate the exposure to chemical carcinogens. However, data on the effect of co-exposure with other chemicals on hemoglobin adduct formation are seldom provided. The objective of this study is to evaluate the effects of pretreatment of ethanol(EtOH) and phenobarbital(PB), which are known to affect metabolism of xenobiotics, on the formation of hemoglobin adducts in the rats(Sprague-Dawley) administered benzidine(BZ). The experimental rats were divided into control, EtOH, and P8 groups. Rats were pretreated with EtOH or PB 24 hours before the oral administration of BZ. Blood sampling was taken before the administration of the chemicals and 0.5, 3, 6, 9, 12, 24, 48, 72, 96, and 144 hours after the administration of the BZ in 5 rats each. The blood was separated into hemoglobin and plasma immediately after taking the blood samples, and the adducts were undergone basic hydrolysis to convert them into aromatic amines. Hydrolyzed BZ, monoacetylbenzidine (MABZ), and 4-aminobiphenyl(4ABP) were separated by reversed-phase liquid chromatography without derivatization, and quantitative analyses of them were performed by a highperformance liquid chromatograph equipped with electrochemical detector. The quantitative amount of the metabolites was expressed by hemoglobin binding index(HBI), BZ-, MABZ-, and 4ABP-HBI of EtOH and PB groups were increased more than those of control group. These results are attributable to the fact that EtOH and PB induced N-hydroxylation related to the hemoglobin adduct formation. The ratio of N-acetylation (viz, MABZ-HBI/BZ-HBI) showed no significant difference between EtOH group and control group. It means that EtOH increased N-hydroxylation and N-acetylation in a similar degree. The N-acetylation ratio of PB group was relatively lower than control group because the PB increased N-hydroxylation induction. The N-acetylation ratios of all groups were higher than 1 during the entire experimental period. This result suggests that the effects of EtOH or PB need to be considered in the biochemical monitoring for the assessment of intermittent exposure of benzidine.
Miscibility of poly(4-vinylpyridine) (P4VP) blends with poly(vinyl acetate-co-vinyl alcohol) (VAc-VAL copolymers) was investigated as a function of comonomer composition of VAc-VAL copolymers. Differential scanning calorimetry (DSC) and thermo-optical microscopic (TOM) analysis confirmed that P4VP is miscible with VAc-VAL copolymers containing more than 30 mole% VAL. Fourier transform inflated (FT-IR) spectroscopic analysis revealed that the strong intermolecular hydrongen bonding interaction between the vinylpyridine and VAL hydroxyl group was formed. Theoretical phase diagram was constructed by the calculation using the Association model, a thermodynamic model for hydrogen-bonded polymer blend systems developed by Coleman et al. The calculated theoretical binodal phase diagrams were in good agreement with the experimentally determined cloud point curves.
Kim, Joong-Hark;Hong, Sung-Kil;Hwang, Sung-Jue;Son, Sang-Wook;Choi, Yong-Soon
Korean Journal of Medicinal Crop Science
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v.20
no.2
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pp.108-116
/
2012
In this study, to develop hair growth agents using natural products which has excellent ability to promote hair growth effect and fewer side effect, animal experiment and clinical trials was performed to evaluate hair growth promotion effect of herbal product containing $Rosa$$mutiflora$ roots extracts as a main component (RMHP). 4-week-old male C57BL/6 mice were removed the dorsal hair using thioglycolate, and applied 15% EtOH solution as a negative control, 5% minoxidil as a positive control and RMHP to dorsal skin. In the results of macroscopy and photo-interpretation, RMHP group recorded 100% (+++++) of hair growth was proved to significantly stimulate hair growth against 80% (++++) negative control group. 37 patients were treated with RMHP and evaluated the therapeutic effect at 16 weeks. Hair density was significantly increased at 16 weeks after applying RMHP ($125.0{\pm}4.9\;FU/cm^2$) compared to before treatment ($104.3{\pm}4.7\;FU/cm^2$, p < 0.05), and hair thickness were also significantly increased ($0.066{\pm}0.003$ mm) than before treatment ($0.055{\pm}0.002$ mm, p < 0.05). The result of clinical photo-interpretation using 7-point rating scale assessment, after 16 weeks clinical symptoms were evaluated to significantly improve with $1.23{\pm}0.05$ (p < 0.05). Therefore, the results of this study were observed that RMHP have hair loss prevention effect and hair growth promotion effect to hair loss patients.
Suhr, Jinhyung;Lee, Hansol;Kim, Suhwan;Lee, Sung Jin;Bae, Eun Young;Ly, Sun Yung
Journal of Nutrition and Health
/
v.55
no.1
/
pp.59-69
/
2022
Purpose: Natural medicinal plant extracts have recently attracted attention as health beneficial foods and potential therapeutic agents for prevention of various diseases. This study was undertaken to measure the anti-inflammatory effect of the ethanol-water fraction obtained from the above-ground portion of Spiraea prunifolia var. simpliciflora, a wild-growing plant in Korea. The final fraction used in this study was the H2O-EtOH (40:60) fraction (SP60), which had the highest antioxidant activity, as determined in previous studies. Methods: The amounts of nitric oxide (NO), tumor necrosis factor (TNF)-α, interleukin (IL)-6 and IL-1β production were measured in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells exposed to SP60. Western blot was performed to measure the expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2, and the activation of nuclear factor (NF)-κB. Results: SP60 exerted no cytotoxicity up to concentrations of 125 ㎍/mL. The levels of inflammatory cytokines, such as NO, TNF-α, IL-6, and IL-1β, were significantly decreased in LPS-stimulated RAW264.7 cells exposed to SP60. In addition, the expression levels of iNOS, COX-2, and phosphorylated p65 showed a concentration-dependent decrease subsequent to SP60 treatment. These results indicate that SP60 inhibits the LPS-induced production of inflammatory cytokines, iNOS, and COX-2, by inhibiting the activation of NF-κB, which is responsible for the expression of inflammatory mediators. Conclusion: The results presented in this study indicate that the H2O-EtOH (40:60) fraction (SP60) extracted from the above-ground portion of Spiraea prunifolia var. simpliciflora has the potential to be developed as a medicine or healthcare food and functional material possessing anti-inflammatory effects. However, it is necessary to first confirm the anti-inflammatory effects of SP60 in in vivo models.
Somok, the heart wood of Caesalpinia sappan is used in traditional Chinese medicine. This study was performed to investigate the effect of growth and culture conditions of brazilin from C. sappan against S. mutans ATCC 25175. The bacteria were cultured in brain heart infusion (BHI) broth, and then incubated under 5% $CO_2$ at $37^{\circ}C$ for 18-24 hours. The effect of brazilin against S. mutans was confirmed under the changes of the culture conditions, such as growth curve and the change of pH, protein, and total carbohydrate. The growth of S. mutans in control medium was the highest at 24 hr, while brazilin-added medium (0.3 mg/ml) showed maximum growth at 32 hr. The pH values of the control medium was 5.25 at 16 hr, but the media supplemented with brazilin (0.3 mg/ml) was 7.0 at 16 hr. The amounts of total carbohydrate of the control medium was 11 mg/ml at 8 hr, but the brazilin-added media (0.3 mg/ml) was 18 mg/ml at 8 hr. In the protein change of the culture medium, the control culture broth and the brazilin supplemented-cultures was 2.4 mg/ml and 2.54 mg/ml at 24 hr, respectively. Polysaccharide contents of the control medium and test media were 3 mg/ml and 2 mg/ml at 8 hr, respectively. Thus, the application of C. sappan can be considered a useful and practical material for the prevention of dental caries.
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