배추의 형질전환용 선발항생제로서 Paromomycin의 이용

조미애,민성란,고석민,유장렬,이준행,최필선,Cho, Mi-Ae,Min, Sung-Ran,Ko, Suck-Min,Liu, Jang-Ryol,Lee, Jun-Haeng,Choi, Pil-Son 한국식물생명공학회 2006 식물생명공학회지 Vol.33 No.4

정상' 배추와 '서울' 배추의 배축절편을 선발마커로서 hygromycin 저항성유전자를 갖고 있는 pCAMBIA1301와 paromomycin저항성유전자를 갖고 있는 pPTN290으로 각각 형질 전환된 LBA4404 또는 EHA101균주와 공동배양한 후 선발배지에서 배양하면서 형질전환체를 선발하였다. 형질전환빈도는 사용된 항생제와 품종에 따라서 현저하게 차이가 있었으며, 특히 paromomycin은 hygromycin보다 효과적이었고 정상 배추는 서울배추보다 양호하였다. 가장 높은 형질전환빈도는 (0.70%) 100mg/L paromomycin이 첨가된 선발배지에서 정상배추의배축을 배양할 경우 얻어졌다. GUS양성반응으로 확인 한 결과 정상배추에서 9개체와 서울배추에서 3개체를 각각 얻었으며, 온실에서 생장한 후 $T_1$종자를 수확하였다. $T_1$ 종자를 다시 발아시켜 유식물체를 얻은 후 GUS양성반응을 확인함으로서 외래유전자가 안정적으로 발현하고 있음을 확인하였다. Hypocotyl explants of Chinese cabbage (us. 'Jeong Sang' and 'Seoul') produced adventitious shoots on Murashige and Skoog (MS) basal medium supplemented with 4mg/L $AgNO_3$, 5 mg/L acetosyringone, 4 mg/L 6-benzyladenine and 3mg/L alpha-naphthaleneacetic acid (SI) after cocoultivation with strains of Agrobacterium tumefaciens (LBA4404) harboring the pCAMBIA1301 and the $_PPTN290$ containing hygromycin-resistance gene and paromomycin-resistance gene as a selectable marker genes, respectively. There was a significant difference in the frequency of transgenic plants depending on antibiotics and cultivars used. Paromomycin was better than hygromycin, and cultivar 'Jeong-sang' was higher than 'c.v. Seoul' in the frequency of transgenic plants. In particular, the highest frequency (0.70%) of transgenic plants was obtained from selection medium (SI) containing 100mg/L paromomycin in c.v., 'Jeong-sang' GUS positive response were obtained 9 plants and 3 plants from the cultivars, 'Jeong-sang' and 'Seoul', respectively. They were grown to maturity in a greenhouse and normally produced $T_1$ seeds. GUS histochemical assay for progeny $(T_1)$ revealed that the transgenes were expressed in the plant genome.

배추의 배축절편으로부터 캘러스와 뿌리 발생을 통한 안정적 형질전환

조미애,김춘해,민성란,고석민,유장렬,최필선,Cho, Mi-Ae,Kim, Choon-Ae,Min, Sung-Ran,Ko, Suck-Min,Liu, Jang-Ryol,Choi, Pil-Son 한국식물생명공학회 2007 식물생명공학회지 Vol.34 No.2

'정상' 배추의 배축절편을 선발마커로서 paromomycin 저항 성유전자를 갖고 있는 pPTN290으로 각각 형질전환된 EHA101, LBA4404, GV3101균주와 공동배양한 후 갤러스유도배지에서 형질전환캘러스를 얻은 후, 뿌리유도배지에서 부정근을 그리고 신초유도배지에서 신초를 각각 순차적으로 유도하였다. 형질전환캘러스를 얻은 후, 뿌리유도배지에서 부정근을 그리고 신초유도배지에서 신초를 가각 순차적으로 유도하였다. 형질전환캘러스 형성은 Agrobacterium균주에 따라 차이가 있었으며, 특히 EHA101균주에 공동배양된 배축절편으로부터 최대 6.1%까지 얻어졌다. 또한 각각의 형질전환캘러스 클론으로부터 형질전환 부정근과 신초 발생은 EHA101균주에서 60.7%와 38.2%, LBA4404에서 8.3%와 0%, GV3101에서 20.5%와 85.7%까지 각각 얻을 수 있었다. 형질전환식물체는 특별한 형태적 이상 없이 온실에서 정상적으로 자라 $T_{2}$종자를 얻을 수 있었다. GUS방법으로 7개의 후대 유식물체를 분석한 결과 gus유전자가 안정적으로 발현하고 있음을 확인하였고, 배추 genome에 single 또는 multiple copy로 전달되고 있음을 추측할 수 있었다. Hypocotyl explants of Chinese cabbage (cvs. "Jeong Sang") produced transgenic calli on callus induction medium (MS salt, B5 vitamin, 5 mg/L acetosyringone, 1 mg/L 2,4-D, 3% sucrose, 400 mg/L cefotaxime, 100 mg/L paromomycin, pH 5.8) after cocultivation with strains of Agrobacterium tumefaciens (EHA101, LBA4404, GV3101) harboring the pPTN290 containing paromomycin-resistance gene as a selectable marker, and then they transferred to root induction medium (1/2MS salt, MS vitamins, 2% sucrose, 100 mg/L paromomycin, 100 mg/L cefotaxime, pH 5.8) and shoot induction medium (MS salt, B5 vitamin, 4 mg/L $AgNO_3$, 4 mg/L 6-benzyladenine, 3 mg/L alpha-naphthaleneacetic acid, 100 mg/L paromomycin, 100 mg/L cefotaxime, 3% sucrose, pH 5.8) in order. There was a significant difference in the frequency of transgenic calli depending on Agrobacterium strains. In particular, the highest frequency (6.1%) of transgenic calli was obtained from the hypocotyls cocultivated with EHA101 strains. Also, the frequency (%) of transgenic root and plants from each transgenic callus clone were obtained with 60.7% and 38.2% in EHA101, with 8.3% and 0% in LBA4404, with 20.5% and 85.7% in GV3101 strains, respectively. They were grown to maturity in a greenhouse and normally produced $T_2$ seeds. GUS histochemical assay for progeny ($T_2$) revealed that the transgenes was expressed in the plant genome, and progeny analysis from 7 independent transgenic events demonstrated that the transformants transmitted the transgene as a single or multiple functional locus.

소동물 폐종양의 정량적 개선을 위한 내부 움직임 평가

유정우,우상근,이용진,김경민,김진수,이교철,박상준,유란지,강주현,지영훈,정용현,김병일,임상무,Yu, Jung-Woo,Woo, Sang-Keun,Lee, Yong-Jin,Kim, Kyeong-Min,Kim, Jin-Su,Lee, Kyo-Chul,Park, Sang-Jun,Yu, Ran-Ji,Kang, Joo-Hyun,Ji, Young-Hoon,Chung, Y 한국의학물리학회 2011 의학물리 Vol.22 No.3

이 연구에서는 폐종양의 정량적 개선을 위하여 분자체를 이용하여 내부 움직임을 측정하고 평가된 데이터를 기반으로 소동물 PET 영상내의 폐종양을 국소화하고자 하였다. 소동물 폐 영역의 내부 움직임은 방사성물질을 흡착한 분자체를 이용하여 소동물 폐 영역에 부착함으로써 구현하였다. 폐 영역의 내부 움직임 표적으로 사용된 분자체는 약 37 kBq의 Cu-64를 흡착시켜 폐종양을 모사하였다. 소동물 PET 영상은 Siemens Inveon 스캐너를 이용하여 획득하였으며 외부 움직임 데이터는 트리거 생성 장치인 BioVet을 이용하였다. SD-Rat PET 영상은 $^{18}F$-FDG 37 MBq/0.2 mL을 미정맥으로 주사하고 60분 후 20분간 데이터를 획득하였다. 리스트모드 데이터의 각 선응답은 외부 트리거 장치에 의해 획득된 트리거신호를 이용하여 2 bin에서 16 bin으로 사이노그램을 획득하였다. 획득된 사이노그램 데이터는 OSEM 2D 알고리즘을 이용하여 4회의 반복으로 재구성하였다. 종양의 정량적 분석을 위한 PET 영상은 종양을 묘사한 분자체 영역에 관심영역을 설정하고 계수와 SNR 그리고 FWHM을 이용하여 평가하였다. 움직임 표적으로 사용된 분자체의 크기는 $1.59{\times}2.50mm$이었으며, 기준 영상으로 획득한 체외 분자체 수직 및 수평 FWHM은 $2.91{\times}1.43mm$이었다. 정적영상과 4 bin 그리고 8 bin 영상에서의 수직 FWHM은 각각 3.90 mm, 3.74 mm, 3.16 mm이었으며 수평 FWHM은 각각 2.21 mm, 2.06 mm, 1.60 mm이었다. 정적영상, 4 bin, 8 bin, 12 bin 그리고 16 bin의 계수 값은 각각 4.10, 4.83, 5.59, 5.38, 5.31이었다. 정적영상, 4 bin, 8 bin, 12 bin 그리고 16 bin의 SNR은 4.18, 4.05, 4.22, 3.89, 3.58이었다. FWHM은 게이트 수의 증가에 따라 계속 향상됨을 확인하였다. 그러나 계수 값과 SNR은 게이트 수의 증가에 따라 계속 향상되지 않고 특정 bin 수에서 가장 높은 값을 보여 소동물 폐 영역에서의 종양 영상화시 SNR의 손실을 최소화하면서 향상된 계수 값을 얻을 수 있는 게이트 수를 획득하였다. 내부 움직임 측정은 최적화된 종양 국소화 영상을 획득할 수 있으며 외부 움직임 모니터링 시스템을 사용하지 않고 장기별 움직임 예측 모델링을 위한 유용한 방법이 될 것으로 기대된다. The purpose of this study was to estimate internal motion using molecular sieve for quantitative improvement of lung tumor and to localize lung tumor in the small animal PET image by evaluated data. Internal motion has been demonstrated in small animal lung region by molecular sieve contained radioactive substance. Molecular sieve for internal lung motion target was contained approximately 37 kBq Cu-64. The small animal PET images were obtained from Siemens Inveon scanner using external trigger system (BioVet). SD-Rat PET images were obtained at 60 min post injection of FDG 37 MBq/0.2 mL via tail vein for 20 min. Each line of response in the list-mode data was converted to sinogram gated frames (2~16 bin) by trigger signal obtained from BioVet. The sinogram data was reconstructed using OSEM 2D with 4 iterations. PET images were evaluated with count, SNR, FWHM from ROI drawn in the target region for quantitative tumor analysis. The size of molecular sieve motion target was $1.59{\times}2.50mm$. The reference motion target FWHM of vertical and horizontal was 2.91 mm and 1.43 mm, respectively. The vertical FWHM of static, 4 bin and 8 bin was 3.90 mm, 3.74 mm, and 3.16 mm, respectively. The horizontal FWHM of static, 4 bin and 8 bin was 2.21 mm, 2.06 mm, and 1.60 mm, respectively. Count of static, 4 bin, 8 bin, 12 bin and 16 bin was 4.10, 4.83, 5.59, 5.38, and 5.31, respectively. The SNR of static, 4 bin, 8 bin, 12 bin and 16 bin was 4.18, 4.05, 4.22, 3.89, and 3.58, respectively. The FWHM were improved in accordance with gate number increase. The count and SNR were not proportionately improve with gate number, but shown the highest value in specific bin number. We measured the optimal gate number what minimize the SNR loss and gain improved count when imaging lung tumor in small animal. The internal motion estimation provide localized tumor image and will be a useful method for organ motion prediction modeling without external motion monitoring system.

Clear cell carcinomas of the ovary: a multi-institutional study of 129 cases in Korea with prognostic significance of Emi1 and Galectin-3.

Min, Kyueng-Whan,Park, Moon Hyang,Hong, Sung Ran,Lee, Heejung,Kwon, Sun Young,Hong, Sook Hee,Joo, Hee Jae,Park, In Ae,An, Hee Jung,Suh, Kwang Sun,Oh, Hoon Kyu,Yoo, Chong Woo,Kim, Mi Jin,Chang, Hee Kyu Raven Press 2013 International journal of gynecological pathology Vol.32 No.1

<P>Accurate diagnosis of ovarian clear cell carcinoma (CCC) is important because of its poor prognosis with chemoresistance and a high recurrent rate. The clinicopathologic characteristics and prognostic significance of the cell cycle regulator [early mitotic inhibitor-1 (Emi1)] and galactoside-binding protein (Galectin-3) were evaluated. Among 155 CCCs from 18 hospitals in Korea between 1995 and 2006, 129 pure CCCs were selected with consensus using immunohistochemical stains for hepatocyte nuclear factor-1β, Wilms' tumor protein, and estrogen receptor. The expressions of Emi1, Galectin-3, p53, and Ki-67 labeling index were analyzed with clinicopathologic parameters and the patient's survival. The mean age of the patients was 49.6 yr; the tumors were bilateral in 10.9%, and the average size was 12 cm. Adenofibromatous component was found in 7%, and endometriosis in 48.1% of the cases. Psammoma body was seen in 16.3%. Disease-free survival and overall survival rates were 78.3% and 79.1%, respectively. The International Federation of Obstetrics and Gynecology (FIGO) stage was the most important prognostic indicator. Emi1 expression (>5%) was seen in 23.3% of CCCs, and associated with high FIGO grades and poor overall survival (P<0.05). High Galectin-3 (80%) expression was seen in 59.7% of CCCs, and associated with FIGO stages III and IV, and high Ki-67 labeling index. High Ki-67 labeling index (50%) and p53 expression (50%) were seen in 27.1% and 18.6% of CCCs, respectively, but there was no clinicopathologic and prognostic significance. On the basis of the fact that the expression of Emi1 in CCC was correlated with a high histologic grade and worse overall survival, target therapy using inhibitors of Emi1 may be tried in the management of CCC patients with Emi1 expression.</P>

Ectoparasiticide efficacy of Derris Extract against Dermanyssus gallinae inhabiting in poultry in larval packet test

Ran Jo,Jeong-Ran Min,Woo-Seog Jeong,Young-Wook Kim,Soo-Hee Kim,Sang-Hee Jeong 한국농약과학회 2020 한국농약과학회 학술발표대회 논문집 Vol.2020 No.11

Association between treatments of psoriasis with severe infectious diseases and malignancies : a nationwide cohort study in Korea

( Sung Min Kim ),( Young Ah Cho ),( Ji Su Lee ),( Ji Youn Hong ),( Jin Hee Kim ),( Joo Ran Hong ),( Hye In Cheon ),( Yang Won Lee ),( Yong Beom Choe ),( Kyu Joong Ahn ) 대한피부과학회 2020 대한피부과학회 학술발표대회집 Vol.72 No.1

Background: There are various treatment options of psoriasis, including phototherapy, systemic oral agents and, recently, biologics. These therapies include immune regulation, so it is necessary to confirm the risk of malignancy and infections. Objectives: To evaluate the risk of malignancies and hospitalized infectious disease (HID) in patients with psoriasis by treatments, comparing with general population (GP). Methods: The study population consisted of 159,423 patients for malignancies and 272,475 patients for HID during 2005 to 2018 from the National Health Insurance Service (NHIS). Adjusted hazard ratios (aHRs) and 95% confidence intervals (CIs) of mild (ML), phototherapy (PH), oral systemics (OR) and biologics (BI) cohort were evaluated. Results: For HID, the results (aHRs; CIs) were ML (1.12; 1.11-1.14), PH (1.05; 1.00-1.09), OR (1.19; 1.15-1.24) and BI (0.85; 0.62-1.17). For tuberculosis (Tb), a particular infectious disease, the results (aHRs; CIs) were ML (1.11; 1.06-1.16), PH (1.11; 0.96-1.28), OR (1.21; 1.08-1.36) and BI (8.83; 6.62-12.5). For malignancies, the results (aHRs; CIs) were ML (1.12; 1.10-1.15), PH (1.20; 1.12-1.29), OR (1.15; 1.08-1.21) and BI (1.47; 0.93-2.43). Conclusion: Patients with psoriasis showed a significantly higher risk of malignancies and HID than GP. Biologics were not seemed to be a significant risk factor for these diseases, but appeared to increase the risk of Tb. Therefore, in choosing a treatment for psoriasis, these risks should be considered.

Analysis of Soil Microbial Communities Formed by Different Upland Fields in Gyeongnam Province

Min Keun Kim,Yong Sik Ok,Jae-Young Heo,Si-Lim Choi,Sang-Dae Lee,Hyun-Yul Shin,Je-Hong Kim,Hye Ran Kim,Young Han Lee 한국토양비료학회 2014 한국토양비료학회지 Vol.47 No.2

The present study investigated variations in soil microbial communities by fatty acid methyl ester (FAME) and the chemical properties at 24 sites of upland soils in Gyeongnam Province. The electrical conductivity of the soil under potato cultivation was significantly higher than those of the red pepper and soybean soils (p < 0.05). The gram-negative bacteria community in potato soil was significantly lower than those in the garlic and soybean soils (p < 0.05). The communities of actinomycetes and arbuscular mycorrhizal fungi in the red pepper soil were significantly higher than those in the potato soil (p < 0.05). In addition, the cy17:0 to 16:1ω7c ratio was significantly lower in red pepper, soybean, and garlic soils compared with potato soil, indicating that microbial stress decreased. Consequently, differences in soil microbial community were highly associated with cultivated crop species, and this might be resulted from the difference in soil chemical properties.

Neonatal outcomes of twin pregnancies delivered at late-preterm versus term gestation based on chorionicity and indication for delivery

Sung, Ji-Hee,Kim, Soo Hyun,Kim, Yoo-Min,Kim, Ji-Hye,Kim, Mi-Na,Lee, Hye Ran,Lee, Hye-Jung,Lee, Eun-Jung,Choi, Suk-Joo,Oh, Soo-young,Roh, Cheong-Rae,Kim, Jong-Hwa De Gruyter 2016 Journal of perinatal medicine Vol.44 No.8

<P><B>Abstract</B></P>Objective: To investigate the neonatal outcomes of twin pregnancies delivered at late-preterm versus term gestation based on chorionicity and indication for delivery.Study Design: This is a retrospective cohort study of women with twin pregnancies delivered at ≥34 weeks of gestation from 1995 to 2014. Subjects were categorized into two groups according to gestational age at delivery: late-preterm group (34–36 weeks) and term group (≥37 weeks). Neonatal outcome measures including neonatal intensive care unit (NICU) admission, mechanical ventilator support, and respiratory distress syndrome (RDS) were compared between the late-preterm and term group based on chorionicity (monochorionic or dichorionic) and delivery indication (elective or non-elective).Results: A total of 1198 twin pregnancies were included in the study: 679 in the late-preterm group and 519 in the term group. Late-preterm twin infants had higher rates of NICU admission, mechanical ventilator support, and RDS than did term twin infants, regardless of the chorionicity and indication for delivery. In the multivariable analysis, late-preterm birth, monochorionicity, and non-elective delivery were independently associated with a significantly higher risk of NICU admission and mechanical ventilator support.Conclusion: The late-preterm birth was associated with a higher risk of adverse neonatal outcome regardless of chorionicity and indication for delivery, and showed significantly increased risk by monochorionicity and non-elective delivery.

Protective Immunity of 56-kDa Type-Specific Antigen of Orientia tsutsugamushi Causing Scrub Typhus

( Sang Ho Choi ),( Hang Jin Jeong ),( Young Ran Ju ),( Byoungchul Gill ),( Kyu Jam Hwang ),( Jeong Min Lee ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.12

Scrub typhus, caused by infection with Orientia tsutsugamushi, is a mite-borne zoonotic disease endemic to the Asian-Pacific region. In Korea, the incidence of this disease has increased with climate changes, and over 10,000 cases of infection were reported in 2013. Although this infection is treatable with antibiotics such as doxycycline and azithromycin, an effective prophylactic vaccine against O. tsutsugamushi would be more desirable for preventing scrub typhus in endemic areas. In this study, we investigated the 56-kDa type-specific antigen (TSA56), which is a major outer membrane protein of O. tsutsugamushi, as a vaccine candidate. Intranasal immunization of recombinant TSA56 (rec56) induced a higher level of TSA56- specific IgG than that induced by intramuscular immunization of tsa56-expressing DNA (p56). Both types of immunization induced a cell-mediated immune response to TSA56, as demonstrated by the splenic cell proliferation assay. Mice immunized with p56, followed by rec56 plus heat-labile enterotoxin B subunit from E. coli, had a stronger protection from a homologous challenge with the O. tsutsugamushi Boryong strain than with other combinations. Our preliminary results suggest that an effective human vaccine for scrub typhus can include either recombinant TSA56 protein or tsa56-expressing DNA, and provide the basis for further studies to optimize vaccine performance using additional antigens or different adjuvants.

Tar colors in medicine: Method validation and screening impurity by High-performance Liquid Chromatography - Photodiode Array Detector

Min Kyeong Kim,Kyeung Ran Min,Do Hui Lee,Hyo Cheon Lee,Geon U Kim,Jong Gi Hong,Yong Seok Choi,Sang Beom Han,Yong Moon Lee 한국분석과학회 2020 학술대회논문집 Vol.2020 No.6