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A Comparative Study between Microbial Fermentation and Non-Fermentation on Biological Activities of Medicinal Plants, with Emphasis on Enteric Methane Reduction (천연 약용식물의 미생물 발효를 통한 장내 메탄 생성 억제 효과 비교 연구)

  • Lee, A-Leum;Park, Hae-Ryoung;Kim, Mi-So;Cho, Sangbuem;Choi, Nag-Jin
    • Korean Journal of Organic Agriculture
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    • v.22 no.4
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    • pp.801-813
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    • 2014
  • A study was conducted to improve the biological activity of two medicinal plants, Eucommia ulmoides Oliv. and Glycyrrhiza uralensis, by fermentation. The biological activity was assessed by determining antibacterial, antioxidant and antimethanogenic properties. Fermentation was achieved by adding the plant materials in MRS broth at 10% (w/v) and different starter cultures at 1% (v/v). Condition for fermentation were incubation temperature of $30^{\circ}C$ and agitation at 150 rpm for 48 h. Six starter cultures, Weissella confusa NJ28 (Genbank accession number KJ914897), Weissella cibaria NJ33 (Genbank accession number KJ914898), Lactobacillus curvatus NJ40 (Genbank accession number KJ914899), Lactobacillus brevis NJ42 (Genbank accession number KJ914900), Lactobacillus plantarum NJ45 (Genbank accession number KJ914901) and Lactobacillus sakei NJ48 (Genbank accession number KJ914902) were used. Antibacterial activity was observed in L. curvatus NJ40 and L. plantarum NJ45 only as opposed to other treatments, including the non-fermented groups, which showed no antibacterial activity. Both plants showed antioxidant activity, although E. ulmoides Oliv. had lower activity than G. uralensis. However, fermentation by all strains significantly improved (p<0.05), antioxidant activity in both plants compared to non-fermented treatment. Six treatments were based on antibacterial activity results, selected for in vitro rumen fermentation; 1) non-fermented E. ulmoides, 2) fermented E. ulmoides NJ40, 3) fermented E. ulmoides NJ45, 4) non-fermented G. uralensis, 5) fermented G. uralensis NJ40, 6) fermented G. uralensis NJ45. A negative control was also added, making a total of 7 treatments for the in vitro experiment. Medicinal plant-based treatments significantly improved (p<0.05) total volatile fatty acid (VFA) concentration. Significant methane reduction per mol of VFA were observed in G. uralensis (p<0.05). Based on the present study, fermentation improves the biological activity of E. ulmoides Oliv. and G. uralensis. Fermented G. uralensis could also be applied as an enteric methane mitigating agent in ruminant animals.

Determinations of Toltrazuril and Toltrazuril Sulfone Levels in Olive Flounder Paralichthys olivaceus Samples Using Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry (LC-MS/MS를 이용한 넙치(Paralichthys olivaceus)시료의 톨트라주릴 및 톨트라주릴 설폰 분석)

  • Hong, Do Hee;Kim, Ah Hyun;Lee, Ka Jeong;Yoon, Minchul;Son, Kwang Tae;Kim, Myoung Sug;Kim, Na Young;Jung, Sung Hee;Jo, Mi Ra
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.52 no.5
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    • pp.461-467
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    • 2019
  • Several studies investigating the prevention and treatment of external parasites in farmed olive flounder Paralichthys olivaceus have found that the anticoccidial agent toltrazuril sulfone is an effective antiparasitic. Prior to undertaking a full-scale study, we developed analytical methods to detect the levels of toltrazuril and toltrazuril sulfone in farmed flounder samples using liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS/MS). This analysis showed that LC-MS/MS changed the mobile phase and collision energy of toltrazuril and toltrazuril sulfone. This was validated using established conditions. Sample pre-treatment for this process involved extraction with dichloromethane and purification by liquid-liquid extraction in formic acid, acetonitrile, and h-hexane, followed by determination of all compounds by LC-MS/MS. Separation was achieved within 10 min by gradient elution using a Capcell Pak C18 ($3.0{\mu}m$, $100{\times}2.0mm$) analytical column (Shiseido UG 120V) with a mixture of 0.1% (v/v) formic acid and acetonitrile. Multiple reaction monitoring was used for selective detection of toltrazuril and toltrazuril sulfone. This method yields satisfactory results for linearity, precision, and limits of quantification. Therefore, the method established in our study will serve as a basis for further research on parasite control by toltrazuril and toltrazuril sulfone.

Optical Properties of SiNx Thin Films Grown by PECVD at 200℃ (200℃의 저온에서 PECVD 기법으로 성장한 SiNx 박막의 열처리에 따른 광학적 특성 변화 규명)

  • Lee, Kyung-Su;Kim, Eun-Kyeom;Son, Dae-Ho;Kim, Jeong-Ho;Yim, Tae-Kyung;An, Seung-Man;Park, Kyoung-Wan
    • Journal of the Korean Vacuum Society
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    • v.20 no.1
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    • pp.42-49
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    • 2011
  • We deposited $SiN_x$ thin films by using PECVD technique at $200^{\circ}C$ with various flow ratios of the $SiH_4/N_2$ gases. The photoluminescence measurements revealed that the maximum emission wavelength shifted to long wavelength as the ratio increased, however, positions of the several peak wavelengths, such as 1.9, 2.2, 2.4, and 3.1 eV, were independent on the ratio. Changes of the photoluminescence spectra were measured in the $N_{2}-$, $H_{2}-$, and $O_2$-annealed films. The luminescence intensities increased after the annealing process. In particular, the maximum emission wavelength shifted to short wavelength after $H_{2}-$ or $O_2$-annealing. But there were still several peaks on the spectra of all annealed films, several peak positions remained to be unchanged after the annealing. As for the light emission mechanism, we have considered the defect states of the Si- and N- dangling bonds in the $SiN_x$ energy gap, so that the energy transitions from/to the conduction/valence bands and the defect states in the gap were attributed to the light emission in the $SiN_x$ films. The experimental results point to the possibility of a Si-based light emission materials for flexible Si-based electro-optic devices.

Spatially-resolved Photoluminescence Studies on Intermixing Effect of InGaAs Quantum Dot Structures Formed by AlAs Wet Oxidation and Thermal Annealing (AlAs 습식산화와 열처리로 인한 InGaAs 양자점 레이저 구조의 Intermixing효과에 관한 공간 분해 광학적 특성)

  • Hwang J.S.;Kwon B.J.;Kwack H.S.;Choi J.W.;Choi Y.H.;Cho N.K.;Cheon H.S.;Cho W.C.;Song J.D.;Choi W.J.;Lee J.I.
    • Journal of the Korean Vacuum Society
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    • v.15 no.2
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    • pp.201-208
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    • 2006
  • Optical characteristics of InGaAs quantum dot (QD) laser structures with an Al native oxide (AlOx) layer as a current-blocking layer were studied by means of photoluminescence (PL), PL excitation, and spatially-resolved micro-PL techniques. The InGaAs QD samples were first grown by molecular-beam epitaxy (MBE), and then prepared by wet oxidation and thermal annealing techniques. For the InGaAs QD structures treated by the wet oxidation and thermal annealing processes, a broad PL emission due to the intermixing effect of the AlOx layer was observed at PL emission energy higher than that of the non-intermixed region. We observed a dominant InGaAs QD emission at about 1.1 eV in the non-oxide AlAs region, while InGaAs QD-related emissions at about 1.16 eV and $1.18{\sim}1.20eV$ were observed for the AlOx and the SiNx regions, respectively. We conclude that the intermixing effect of the InGaAs QD region under an AlOx layer is stronger than that of the InGaAs QD region under a non-oxided AlAs layer.

tans-Resveratrol Content of Varieties and Growth Period in Peanut (땅콩 품종 및 생육기별 trans-Resveratrol 함량)

  • Lee, Mi-Ja;Cheong, Young-Keun;Kim, Hyung-Soon;Park, Ki-Hun;Doo, Hong-Soo;Suh, Duck-Yong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.6
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    • pp.429-433
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    • 2003
  • trans-Resveratrol(3,5,4'-trihydroxystilbene) is phenolic compound present in grapes, wines, and peanuts, has been reported to have health benefits including anticarcinogenic effects, protection against cardiovascular diseases and reduced cancer risk. A simple method for the quantitative extraction of trans-resveratrol from peanut has been developed. Optimal conditions for extraction were investigated. Type of solvent, time, and temperature assayed influenced trans-resveratrol yield. Adequate extraction condition was decided to ethanol/water (80:20v/v) maintained at $25^{\circ}C$ for 45 min. After extraction, the protocol consists of sample preparation using a $\textrm{C}_{18}$ solid-phase extraction (SPE) cartridge after concentrate with rotary evaporator and quantified by reversed phase HPLC using a $\textrm{C}_{18}$ column at 308 nm. Analytical methods for measuring trans-resveratrol in peanut were adapted to isolate, identify, and quantify trans-resveratrol in 11 peanut varieties by HPLC (high-performance liquid chromatography) with UV detector, The 11 peanut varieties content ranged from 0.018 to 1.125 $\mu\textrm{g}/\textrm{g}$ with an average of 0.289 $\mu\textrm{g}/\textrm{g}$. The contents were higher in the seeds with than without testa, regardless of varieties. The trans-resveratrol content was Higher in 110, 130 days after sowing than that of other period.

Results of Radiotherapy in Nasopharyngeal Cancer (비인두암의 방사선치료 결과)

  • Shin Byung Chul;Ma Sun Young;Moon Chang Woo;Yum Ha Yong;Jeung Tae Sig;Yoo Myung Jin
    • Radiation Oncology Journal
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    • v.13 no.3
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    • pp.215-223
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    • 1995
  • Purpose : The aim of this study was to assess the effectiveness, survival rate and complication of radiation in nasopharyngeal cancer. Materials and Methods : From January 1980 to May 1989. Fifty patients who had nasopharyngeal carcinoma treated with curative radiation therapy at Kosin Medical Center were retrospectively studied. Thirty seven patients($74{\%}$) were treated with radiation therapy alone(Group I) and 13 patients ($26{\%}$) treated with combination of chemotherapy and radiation (Group II). Age distribution was 16-75 years(median : 45.8 years). In histologic type, squamous cell carcinoma was in 30 patients($60{\%}$), undifferentiated carcinoma in 17 patients($34{\%}$), and lymphoepithelioma in 3 patients($6{\%}$). According t AJCC staging system. 4 patients($8{\%}$) were in $T_1$, 13 patients($26{\%}$) in $T_2$. 20 patients($40{\%}$) in $T_3$, 13 patients($26{\%}$) in $T_4$ and 7 patients($14{\%}$) in $N_0$, 6 patients($12{\%}$) $N_1$, 23 patients($46{\%}$) in $N_2$, 14 patients($28{\%}$) in $N_3$. Total radiation dose ranges were 5250-9200cGy(median : 7355 cGy) in Group I and 5360-8400cGy(median : 6758cGy) in Group II Radiotherapy on 4-6MV linear accelerator and/or 6-12MeV electron in boost radiation was given with conventional technique to 26 patients($52{\%}$), with hyperfractionation(115-120cGy/fr., 2times/day) to 16 patients($32{\%}$), with accelerated fractionation(160cGy/fr., 2 times/day) to 8 patients($16{\%}$). In chemotherapy, 5 FU 1000mg daily for 5 consecutive days, pepleomycin 10mg on days 1 and 3, and cisplatin 100mg on day 1 were administered with 3weeks interval, total 1 to 3 cycles(average 1.8cycles) prior to radiation therapy. Follow up duration was 6-140 months(mean : 58 months). Statistics was calculated with Chi-square and Fisher's exact test. Results : Complete local control rates in Group I and II were $75.7{\%},\;69.2{\%} Overall 5 year survival rates in Group I and II were $56.8{\%},\;30.8{\%}$. Five year survival rates by histologic type in Group I and II were $52.2{\%},\;14.3{\%}$ is squamous cell carcinoma and $54.5{\%},\;50{\%}$ in undifferentiated carcinoma. Survival rates in Group I were superior to those of Group II though there were not statistically significant. In both group, survival rates seem to be increased according to increasing total dose of radiation up to 7500cGy, but not increased beyond it. There were not statistically significant differences in survival rates by age, stage, and radiation techniques in both group. Twenty four patients($48{\%}$) experienced treatment failures. Complications were found in 12 patients($24{\%}$). The most common one was osteomyelitis(4 patients, $33.3{\%}$) involving mandible (3 patients) and maxilla(1 patient). Conclusion : Chemotherapy in combination with radiotherapy was found to be not effective to nasopharyngeal cancer and the survival rate was also inferior to that of radiation alone group though it was statistically not significant due to small population in chemotherapy combined group.

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A Novel Tetrameric Assembly Configuration in VV2_1132, a LysR-Type Transcriptional Regulator in Vibrio vulnificus

  • Jang, Yongdae;Choi, Garam;Hong, Seokho;Jo, Inseong;Ahn, Jinsook;Choi, Sang Ho;Ha, Nam-Chul
    • Molecules and Cells
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    • v.41 no.4
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    • pp.301-310
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    • 2018
  • LysR-type transcriptional regulators (LTTRs) contain an N-terminal DNA binding domain (DBD) and a C-terminal regulatory domain (RD). Typically, LTTRs function as homotetramers. VV2_1132 was identified in Vibrio vulnificus as an LTTR that is a homologue of HypT (also known as YjiE or QseD) in Escherichia coli. In this study, we determined the crystal structure of full-length VV2_1132 at a resolution of $2.2{\AA}$, thereby revealing a novel combination of the domains in the tetrameric assembly. Only one DBD dimer in the tetramer can bind to DNA, because the DNA binding motifs of the other DBD dimer are completely buried in the tetrameric assembly. Structural and functional analyses of VV2_1132 suggest that it might not perform the same role as E. coli HypT, indicating that further study is required to elucidate the function of this gene in V. vulnificus. The unique structure of VV2_1132 extends our knowledge of LTTR function and mechanisms of action.

The Production of Chimeric Mice by Embryonic Stem Cell Carrying Human Luteinizing Hormone Gene (사람 LH 호르몬유전자를 도입한 배아주세포에 의한 카이미라 생쥐 생산)

  • ;Tokunaga, T.;Imai, H.
    • Journal of Embryo Transfer
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    • v.9 no.3
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    • pp.243-247
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    • 1994
  • 최근 의약적으로 유용한 단백질을 대량 생산키 위한 실현 가능한 방법이 유전자변환 가축의 이용과 관련되어 발전되어 왔다. 이러한 유전자 변환동물은 이종의 단백질을 유즙속으로 분비시키는 생체반응기로서 이용되고 있다. 이러한 전략적 목적을 위해 현재 유전자 변환동물의 생산을 위한 이용에 있어 여러 가지 방법들이 보고되고 있다. 그러나 ES 세포의 사용이 이러한 방법들 사이에서 가장 실질적인 것으로 추정되고 있다. 본 실험에서는 유전자 구축을 위해 사람 황체 호르몬(human luteinizing hormone; hLH)의 전사를 유도하기 위해 각각 2.2 및 0.5 kb의 토끼 $\beta$-casein pronoter 단편을 이용하여 생쥐의 유선에 hLH를 발현시키도록 조절하고 발현이 thynidine kinase(TK) pronoter에 의해 좌우되는 neo 유전자를 selectable marker로서 plasnid속에 삽입하였다. 그 결과 생긴 구축 유전자는 각각 pCas 2.2와 pCas 0.5로 명명하였다. 구축된 유전자로 2$\times$107의 TT-2 ES세포를 170V, 550$\mu$F로 100$\mu$g의 선상 plasmid에 의해 electroporation 시켰다. 감염된 colony들은 250$\mu$g/$m\ell$ G418을 함유하는 ESM 배양액에서 선별 7일 이후에 회수하여 성공적으로 감염된 ES세포는 PCR 및 Southern blot에 의해 확인되었고 그들 중 나머지는 trypsin 처리 후 각각 미세조작과 공배양 기술을 사용하여 ICR 생쥐의 8세포기 수정란 속에 도입하였다. 결국 24시간 동안 37$^{\circ}C$, 5% $CO_2$에서 배양된 배반포를 chimera의 생산을 위해 위임신 유기된 G418 선발처리 이후 400 및 275개의 ES 세포 colony가 생존하였으며, 3개의 ES 세포으 colony 의 genome 속에 임의적으로 plamid가 삽입된 것을 Southern blot에 의해 확인되었다. 총 13 chimera 생쥐가 3 colony로부터 생산되었으나 germ-line chimera는 현재 조사중이다. chimera 생산빈도는 공배양 기술보다 주입방법에서 현저히 높았다.

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A Comparison of Sampling and Analytical Methods for Airborne Isocyanates (공기중 이소시아네이트류의 측정 및 분석방법에 관한 비교연구)

  • 변혜정;윤충식;백남원
    • Journal of Environmental Health Sciences
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    • v.22 no.2
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    • pp.32-42
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    • 1996
  • This study was performed to evaluate accuracy and precision of filter method and impinger method for analyzin airborne isocyanates in mixture (2, 6-TDI, HDI, 2, 4-TDI, MDI). Filter method was performed using the OSHA Method 42 and impinger method using the NIOSH Method 5521. The samples were analyzed by high performance liquid chromatography-ultraviolet detector (HPLC-UVD). After the optimum operating conditions for each method were investigated, samples with various concentration levels were quantified at the conditions. The precision was expressed by the pooled coefficient of variation(C.V.) and the accuracy by overall accuracy. The results are summarized as follows: 1. The optimum condition of filter method was determined at 35/65 ACN/buffer (0.01 M ammonium acetate) in mobile phase. And in case of impinger method, it was at 30/70 ACN/buffer(0.2 M sodium acetate). The effect of concentrations of acetate on the separation of the peaks was not significant, but, the effect of ACN/buffer ratio was significant. 2. The correlation coefficients for the two methods were above 0.9 in all isocyanate compounds. Average recovery efficiencies for 2, 6-TDI, HDI, 2, 4-TDI and MDI in filter method were 92.4%, 102.6%, 87.3% and 101.0%, respectively. Those in impinger method were 106.6%, 106.7%, 99.0% and 103.6%, respectively. As a result, the recovery efficiency of impinger method was higher than those of filter method in analyzing isocyanate compounds. 3. The pooled coefficients of variations of the methods were slightly higher than expected. The overall accuracies of the methods were within $\pm 25%$ for each isocyanate compound. Since these results satisfy NIOSH criteria, the accuracy of the experiment is appropriate. 4. As seen above, impinger method is more efficient than filter method. But, there are many disadvantages in impinger method. Therefore, solid sorbent such as a glass fiber filter must be developed in order to have the high efficiency not less than that of impinger method in the future.

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Identification of Oligosaccharides in Human Milk Bound onto the Toxin A Carbohydrate Binding Site of Clostridium difficile

  • Nguyen, Thi Thanh Hanh;Kim, Jong Woon;Park, Jun-Seong;Hwang, Kyeong Hwan;Jang, Tae-Su;Kim, Chun-Hyung;Kim, Doman
    • Journal of Microbiology and Biotechnology
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    • v.26 no.4
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    • pp.659-665
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    • 2016
  • The oligosaccharides in human milk constitute a major innate immunological mechanism by which breastfed infants gain protection against infectious diarrhea. Clostridium difficile is the most important cause of nosocomial diarrhea, and the C-terminus of toxin A with its carbohydrate binding site, TcdA-f2, demonstrates specific abolishment of cytotoxicity and receptor binding activity upon diethylpyrocarbonate modification of the histidine residues in TcdA. TcdA-f2 was cloned and expressed in E. coli BL21 (DE3). A human milk oligosaccharide (HMO) mixture displayed binding with TcdA-f2 at 38.2 respond units (RU) at the concentration of 20 μg/ml, whereas the eight purified HMOs showed binding with the carbohydrate binding site of TcdA-f2 at 3.3 to 14 RU depending on their structures via a surface plasma resonance biosensor. Among them, Lacto-N-fucopentaose V (LNFPV) and Lacto-N-neohexaose (LNnH) demonstrated tight binding to TcdA-f2 with docking energy of −9.48 kcal/mol and −12.81 kcal/mol, respectively. It displayed numerous hydrogen bonding and hydrophobic interactions with amino acid residues of TcdA-f2.