• Journal of Species Research
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• 제10권2호
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• pp.154-158
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• 2021

An endemic endangered species, Aster altaicus var. uchiyamae (Danyang aster) B2015-0044, is cultivated at the Shingu Botanical Garden, which serves as the ex situ conservation institution for this species. In this work, we sequenced the chloroplast genome of A. altaicus var. uchiyamae B2015-0044. We found that the chloroplast (cp) genome of B2015-0044 was 152,457 base pairs(bps) in size: 84,247 bps of large single copy regions(LSC), 25,007 bps of inverted repeats(IRs), and 18,196 bps of small single copy regions. The B2015-0044 cp genome contains 79 protein-coding genes (PCGs), 4 RNA genes, 29 tRNA genes, and 3 pseudogenes. These results were identical to a previously reported cp genome (Park et al., 2017), except for two sites in introns and three in intergenic spacer (IGS) regions. For the intronic differences, we found that clpP.i1 had a 1-bp small simple repeat (SSR) (T) and petD.i had a 3-bp SSR (ATT). We found 1-bp SSRs in the IGSs of trnT_ggu~psbD and psbZ~trnG_gcc, C and A, respectively. The IGS of(ndhF)~rpl32 had a SNP. Based on our results, the cp genome of the A. altaicus var. uchiyamae can be classified into two genotypes, [C]¹-[A]¹²-[T]¹²-[ATT]⁴-C and [C]²-[A]¹¹-[T]¹¹-[ATT]²-A.

• Journal of Applied Biological Chemistry
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• 제53권3호
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• pp.132-138
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• 2010

B. pseudomycoides로 부터 alanine racemase로 추정되는 유전자를 분리한 다음 6xHistidine 이 결합된 pET-21 운반체를 이용하여 E. coli BL21(DE3)에서 발현시키고 생화학 특성을 조사하였다. 재조합된 alanine racemase는 affinity chromatography를 이용하여 분리하였으며 SDS-PAGE 분석에서 약 46 kDa의 단일밴드를 나타내었다. 분리된 효소는 여러 아미노산 중에서 L-alanine에 대하여 가장 높은 활성도를 보이고 D-alanine에 대하여 두번째로 높은 활성도를 보였다. 따라서 분리된 효소는 alanine racemase로 판단되었다. 분리된 효소는 D-cysteine에 하여 상당히 저해가 되었다. 효소의 최적 활성도는 pH 9.0 근처에서 관찰되었고 산성의 조건보다는 알칼리의 조건에서 보다 안정하였다. 보효소인 PLP 0.3mM의 첨가에 의해 효소의 활성도는 약 70% 가량 증대되었다.

• Natural Product Sciences
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• 제14권1호
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• pp.37-46
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• 2008

From the 70% EtOH extract of the roots of Astragalus membranaceus (Leguminosae), fifteen saponins were isolated and identified as astragaloside I (1), isoastragaloside II (2), astragaloside II (3), agroastragaloside I (4), cyclogaleginoside B (5), cycloaraloside A (6), brachyoside B (7), agroastragaloside II (8), astragaloside III (9), astragaloside IV (10), astramembranoside A (11), astramembranoside B (12), cylocanthoside E (13), cyclounifolioside B (14) and azukisaponin V methyl ester (15) by spectroscopic methods. Ten compounds 1 - 3, 5 - 7, 9 - 11 and 14 have cycloastragenol as an aglycon, and four compounds 4, 8 , 12, and 13 have cyclocanthogenin as an aglycon. The hairy roots of A. membranaceus were shown to produce previously unreported cycloartane-type saponins such as agroastragalosides I (4) and II (8) and cycloastragenol $3-O-{\beta}-D-xyloside$ (5), together with the known saponins. This is the first report of these saponins (4, 5, and 8) from the intact plant. Although the occurrence of the oleanane-type triterpene saponin, azukisaponin V methyl ester (15), in Astragalus plants has been demonstrated by others, this is the first report of the azukisaponin V methyl ester (15) from the Astragalus plants.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 춘계학술대회
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• pp.33-33
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• 2019

The AKT signaling pathway is a highly regulated cell signaling system that forms a network with other cell signaling pathways. Hence, the AKT signaling pathway mediates several important cellular functions that include cell survival, proliferation, cell migration, and et cetera. Irregularities that led overactive AKT signaling have been linked to many diseases such as cancer and metabolic-associated diseases. Hence, modulating the overactive AKT signaling pathway via inhibitor is a tantalizing prospect for treatment of cancer and metabolic-associated diseases. Two inhibitors of the AKT signaling pathway will be presented in this symposium: 1) Bisleuconothine A (BisA), a bisindole alkaloid that inhibit autophagy and 2) Ceramicine B (CerB), a limonoid that inhibit adipogenesis. The first topic is on a bisindole alkaloid, BisA and its mechanism in inducing autophagosome formation in lung cancer cell line, A549.(1) Since most autophagy inducing agents generally induce apoptosis, we found that BisA does not induce apoptosis even in high dose. BisA up-regulation of LC3 lipidation is achieved through mTOR inactivation. The phosphorylation of PRAS40, a mTOR repressor was suppressed by BisA. This observation suggested that BisA inactivates mTOR via suppression of PRAS40 phosphorylation. Interestingly, the phosphorylation of AKT, an upstream regulator of PRAS40 phosphorylation was also down-regulated by BisA. These findings suggested that Bis-A induces autophagosomes formation by interfering with the AKT-mTOR signaling pathway. The second topic is on CerB and its mechanism in inhibiting adipogenesis in preadipocytes cell line, MC3T3-G2/PA6.(2,3) CerB inhibits the phosphorylation of protein kinase B (AKT) at the Thr308 position but not the Ser473. Consequently, the phosphorylation of FOXO3 which is located downstream of AKT is also inhibited. Considering that FOXO3 is an important regulator of PPARγ which is a key factor in adipogenesis, CerB may inhibit adipogenesis via the AKT-FOXO3 signaling pathway. Taken together, both BisA and CerB highlighted the potential of AKT signaling pathway modulation as an approach to induce autophagy and inhibit the formation of fat cells, respectively.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2022년도 추계학술대회
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• pp.100-100
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• 2022

Hovenia dulcis is a herbal plant, which belongs to the Rhamnaceae family and is a native of Japan, China and Korea. Its fruit stalk is called 'Jiguja' in Korea. It has been traditionally used as a medicinal plant in East Asia. It was reported to have detoxification effects on alcohol poisoning, and antioxidant, antidiabetic etc. Sample of 5 g was extracted with 50 mL of 70% EtOH. The supernatant was filtered by 0.45 ㎛ membrane filter before analysis. The UPLC system was performed on Waters alliance UPLC HSS T3 column (2.1 × 100 mm, 1.7 ㎛) with a UV detector. The gradient system was a binary eluent of 0.1% formic acid in water(A) and 0.1% formic acid in acetonitrile(B) with gradient conditions as follows: Initial, 10% B; 1 min, 10% B; 4 min, 20% B; 10 min, 25% B; 12 min, 30% B; 14 min, 90% B; 17 min, 90% B; flow rate of 0.2 mL/min. The samples were injected by 2 µL and were detected at UV 355 nm. As a result of analysis, chromatographic patterns appeared in two cases: samples analyzed for ampelopsin and myricetin, and samples analyzed for taxifolin and quercetin. Among the four compounds, the largest regional difference was found to be taxifolin.

• Parasites, Hosts and Diseases
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• 제26권2호
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• pp.87-94
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• 1988

Kim et at.(1986)은 유구낭미충 낭액을 감작시켜 만든 단세포군 항체를 CNBr활성화 Sepharose 4B에 연결시켜 친화성 크로마토그래피를 실시하고 낭액에서 A-항원을 분리한 바 있다. 이 연구는 그 항원 단백질의 생화학적 성상을 관찰한 것이다. Disc-PAGE에 의해 4.5~10% 폴리아크릴아마이드 젤에서 나타내는 Rf간을 기초로 분자량을 측정한 결과 A 항원의 분자량은 150,000 dalton이었다. A-항원은 SDS-PAGE에서 15,000, 10,000, 7,000 dalton에 해당하는 polypeptide로 분획이 구별되었다. 10% 2-mcrcaptoethanol로 처리하지 않거나 $95^{\circ}C$에서 5분간 처리하지 않은 A-항원의 SDS-PAGE에서는 7,000 dalton의 polypeptide가 분리되지 않았다. A-항원의 등전점(PI)은 pH6.8이었다. 낭액항원의 각 분획에 반응하는 항체를 갖는 환자혈청으로 A-항원을 SDS-PAGE/EITB한 바 환자혈청은 15,000, 10,000, 7,000 dalton에 해당하는 부위에서만 반응하였다. 낭액 단백질의 약70%를 차지하는 A-항원은 면역학적으로 내열성을 가졌으며 포충(hydatid cyst) 낭액 항원중 Oriol et at.(1971)의 "Antigen B"와 생화학적 성상이 비슷한 단백질이었다.

• Clinical and Experimental Reproductive Medicine
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• 제26권1호
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• pp.75-81
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• 1999

The beneficial effect of glucose and phosphate ions in culture medium on the development of human embryos in vitro has not been fully elucidated. The purpose of this study was to evaluate the influence of fertilization and culture of embryos in glucose/phosphate-free m-TALP medium on pregnancy rates in IVF-ET program. The patients in 244 IVF-ET cycles received GnRH agonist + HMG regimens. A does of 10,000 IU HCG was administered when two or more dominent follicles reached 18mm in diameter. Thirty-six hours after HCG, oocytes were recovered transvaginally using ultrasound guidance. Aspirated oocytes were matured for 4 to 6 h in TCM-199 supplemented with 10% follicular fluid (FF). Insemination was carried out with 50,000 motile spermatozoa in TCM-199 + 10% FF or m-TALP + 5% FF + 5% fetal cord serum (FCS) according to experimental design. After 6 h, oocytes were washed 3 to 4 times and cultured in each fresh medium. After 20 h, oocytes were freed from cumulus/corona cells and examined for the presence of pronuclei. Fertilized oocytes were transferred into each co-culture drops and cultured for further incubation. On day 3, embryo transfer was performed with grade 1 and 2 embryos. Monolayers for co-culture of embryos were prepared by plating $1{\times}10^5$ cumulus cells/ml in 10ul drop of TCM-199 + 10% FF or m-TALP + 5% FF + 5% FCS media 24 h prior to the onset of co-culture. Development to 4 to 16 cell stage was observed at 70x magnification following two days of incubation. Pregnancy was confirmed by detecting increasing serum ${\beta}$-hCG concentrations for 11 days following embryo transfer. Data were analyzed by ${\chi}^2$-test. Oocytes from 244 IVF-ET cycles were randomized. The number of cycles and mean age of patients were 97 and 147, 31.3 yrs and 31.2 yrs for TCM-199 (control) and m-TALP groups, respectively. The mean number of retrieved oocytes/cycle, fertilization rates, number of embryos transferred/ET and pregnancy rates were 11.1 and 10.3, 65.1% and 67.3%, 4.1 and 4.7, 28.9% and 43.8% for TCM-199 and m-TALP groups, respectively. Differences in the pregnancy rates were found between control and m-TALP groups (p<0.05). The pregnancy rate of patients divided according to maternal age groups of ${\leq}30$, 31-35, $36{\leq}$ were 44.4% and 49.0%, 26.1% and 41.3%, 29.2% and 41.2% for control and m-TALP groups, respectively. These data indicate that culture of human embryos in glucose/phosphate-free m-TALP medium improves pregnancy rates.

• Journal of Korean Neurosurgical Society
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• 제58권6호
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• pp.528-533
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• 2015

Objective : We sought to examine anatomic variations of the atlas and the clinical significance of these variations. Methods : We retrospectively reviewed 1029 cervical 3-dimensional (3D) CT images. Cervical 3D CT was performed between November 2011 and August 2014. Arcuate foramina were classified as partial or complete and left and/or right. Occipitalization of the atlas was classified in accordance with criteria specified by Mudaliar et al. Posterior arch defects of the atlas were classified in accordance with criteria specified by Currarino et al. Results : One hundred and eight vertebrae (108/1029, 10.5%) showed an arcuate foramen. Bilateral arcuate foramina were present in 41 of these vertebrae and the remaining 67 arcuate foramina were unilateral (right 31, left 36). Right-side arcuate foramina were partial on 18 sides and complete on 54 sides. Left-side arcuate foramina were partial on 24 sides and complete on 53 sides. One case of atlas assimilation was found. Twelve patients (12/1029, 1.17%) had a defect of the atlantal posterior arch. Nine of these patients (9/1029, 0.87%) had a type A posterior arch defect. We also identified one type B, one type D, and one type E defect. Conclusion : Preoperative diagnosis of occipitalization of the atlas and arcuate foramina using 3D CT is of paramount importance in avoiding neurovascular injury during surgery. It is important to be aware of posterior arch defects of the atlas because they may be misdiagnosed as a fracture.

• 동의생리병리학회지
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• 제26권6호
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• pp.908-914
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• 2012

To investigate the difference and change of ingredient and efficacy of Galgeun-tang (GGT) according to extraction solvent, water and 70% EtOH, the quantities of index components and the several in vitro activities of two kinds of GGT extract were compared. The contents of extracts were analyzed with HPLC. The biological activities such as anti-inflammatory and anti-obesity effects were measured through cell line-based in vitro assay. The cytotoxicity was measured by CCK-8 assay in RAW 264.7 cell, BEAS-2B cell, HaCaT cell and 3T3-L1 cell. We compared effects of two kinds of extract by measuring NO, PGE2, IL-6 and TNF-${\alpha}$ in RAW264.7 cell, RANTES in BEAS-2B cell, MDC and RANTES in HaCaT cell and GPDH activity and leptin level in differentiated 3T3-L1. 70% EtOH extract of GGT contained more compositions than water extract. The inhibitory effect of water extract of GGT on NO in RAW 264.7 cell and GPDH activity in 3T3-L1 was stronger than that of 70% EtOH. 70% EtOH has a stronger inhibitory effect on PGE2 in RAW264.7 cell, RANTES in BEAS-2B cell, MDC, RANTES in HaCaT cell, leptin in 3T3-L1 cell than water extraction. These results suggest that the ingredient and efficacy from 70% EtOH extract of GGT are more effective than water extract.

• 한국버섯학회지
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• 제15권4호
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• pp.183-189
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• 2017

양송이배지로부터 분리한 인산가용화균 B. metallica JH-7과 B. contaminans JH-15의 단일접종 및 동시접종에 따른 인산가용화 능력의 시너지효과 및 상추생육효과를 조사하였다. 인산캄슘이 함유된 NBRIP 액체배지에 분리균을 접종하고 해리된 가용인산(soluble phosphorus)함량을 HPLC에 의해 분석하여 인산가용화능을 측정한 결과, B. metallica $140.08{\mu}g\;mL^{-1}$, B. contaminans $135.95{\mu}g\;mL^{-1}$, 그리고 동시접종 $134.84{\mu}g\;mL^{-1}$ 순으로 나타나 두 종의 인산가용화 박테리아간의 동시접종에 의한 시너지효과가 없었다. 배지 내 pH 와 잔류 glucose 함량 변화도 모든 접종구에서 배양 1일 후 대부분의 변화가 이루어져 pH의 경우 초기 pH 7.0에서 pH 4.5 수준으로 감소하였으며, 잔류 glucose 함량은 초기 $10mg\;mL^{-1}$에서 $5.3mg\;mL^{-1}$ 수준으로 검출되어 그 이후에는 큰 변화를 보이지 않아 인산가용화능과 거의 유사한 경향을 보였다. 배양여액의 유기산 분석결과 succinic acid, glutamic acid, malic acid, lactic acid를 확인할 수 있었으며, Park et al(2016)에 의해 보고된 인산가용화균의 결과와는 달리 gluconic acid 및 oxalic acid는 검출되지 않았다. 유기산 중에서 lactic acid 와 glutamic acid 가 가장 높은 함량인 $0.18mg\;mL^{-1}$과 $0.16mg\;mL^{-1}$을 나타내었으며, succinic acid는 B. metallica 에서만 확인되었고, 대부분의 유기산이 배양 1일 과 3일 후 생성되는 결과를 보임으로서 유기산이 배지의 pH를 감소시키고 인산 가용화를 유도하는 주요 원인임을 확인할 수 있었다. 접종 1주 후와 2주 후 간의 생육차이를 조사한 상추재배실험결과, 잎의 길이는 2.60 cm 로 B. metallica 처리구가 가장 높았고, 전체길이는 B. metallica + TCP 처리구가 5.50 cm로 가장 높았으며, B.contaminans + TCP 처리구는 잎의 폭과 뿌리길이의 변화에서 0.87 cm 와 3.87 cm로 가장 높았다. 또한 잎의 수는 동시접종 + TCP 처리구에서 6.67 cm로 가장 높은 결과를 보여 재배실험에서도 단일접종과 동시접종구간에 유의수준의 차이를 확인할 수 없었으나, 다른 연구와 유사한 대조구와 비교하였을 경우 인산가용화균을 접종한 모든 처리구에서 뚜렷한 생장촉진효과를 나타냄으로서 접종제로서 인산가용화균을 사용하면 식물과 작물 수확량에 의한 P 섭취가 동시에 증가하고 생육이 촉진된다는 다른 연구결과($Rodr\acute{i}guez$ et al, 1999; Walpola and Yoon. 2013)와 유사한 경향을 보였다. 이 결과를 통하여 인(P) 결핍 토양에 가능한 접종 도구로서 효율적인 PSB 균주를 선택하는 것이 중요하며, 좀 더 정확한 결과를 얻기 위해서는 pot 실험 이후 추가적으로 노지 밭과 시설 재배지 토양의 EC, pH, 유기물 등과 칼륨, 칼슘, 마그네슘과 같은 교환 가능한 양이온 등 화학적 분석(Suh et al, 2008) 등과 이를 기초로 하여 토양의 포트 및 면적에 따른 미생물의 적절한 접종량 확인 등이 더 필요 할 것이라고 생각된다.