Slide Session : OS-IFD-07 ; Infectious Disease : In Vitro Antiviral Activity of Ribavirin Against Severe Fever with Thrombocytopenia Syndrome Virus

( Myung Jin Lee ),( Kye Hyung Kim ),( Jong Youn Yi ),( Su Jin Choi ),( Chung Jong Kim ),( Nak Hyun Kim ),( Kyoung Ho Song ),( Pyoeng Gyun Choi ),( Ji Hwan Bang ),( Wan Beom Park ),( Eu Suk Kim ),( San 대한내과학회 2014 대한내과학회 추계학술대회 Vol.2014 No.1

In Vitro Antiviral Activity of Ribavirin Against Severe Fever with Thrombocytopenia Syndrome Virus Myung Jin LEE1, Kye-Hyung KIM1, Jongyoun YI2, SuJin CHOI1, Chung-Jong KIM1, Nak- Hyun KIM1, Kyoung-Ho SONG1, Pyoeng Gyun CHOI1, Ji-Hwan BANG1, Wan Beom PARK1, Eu Suk KIM1, Sang-Won PARK1, Hong Bin KIM1, Nam Joong KIM1, Myoung- Don OH1 Seoul National University College of Medicine, Korea1, Pusan National University School of Medicine, Korea2 Background: Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease caused by a novel Bunyavirus, severe fever with thrombocytopenia syndrome virus (SFTSV). No effective antiviral therapy is proven yet, but clinical use of ribavirin (RBV) has been tried. We investigated the antiviral effect of RBV against SFTSV in vitro. Methods: To test for cytotoxicity of RBV, Vero cells were treated with different concentrations of RBV (3.90 to 500 μg/mL, two-fold dilution) and analyzed by cell viability MTS assay 48h post-infection. To determine antiviral activity of RBV against SFTSV, Vero cells were infected with SFTSV strain Gangwon/Korea/2012 at 100 TCID50 (50% tissue culture infective dose) per well in a 96-well plate, and RBV was added at the concentrations showing no or minimal cytotoxicity. Viral RNAs were extracted from the culture supernatants and quantifi ed using one-step real-time reverse transcription- PCR to amplify the partial large segment of SFTSV. Statistical analysis was done by one-way ANOVA with Tukey`s post hoc test. Results: Cytotoxicity due to RBV was not observed at RBV concentration =31.3 μg/ mL. Viral RNAs at 24h post-RBV treatment were reduced with increasing RBV concentrations (1-32 μg/mL), compared with those of mock-treated cells (P <0.01, Figure). Half maximal inhibitory concentration (IC50) of RBV was 3.69 μg/mL at 24h post-RBV treatment. Conclusions: Our study shows that RBV has antiviral effect against SFTSV in a dose-dependent manner. Further studies are required to evaluate the effi cacy of RBV in SFTS.

Effects of Phenytoin and Diazepam on the Seizure Activity in the Cortical Dysplasia Animal Models

Kim, Si-Hyung,Choi, In-Sun,Cho, Jin-Hwa,Park, Eun-Ju,Jang, Il-Sung,Choi, Byung-Ju,Kim, Hyun-Jung,Kim, Young-Jin,Nam, Soon-Hyeun The Korean Academy of Oral Biology 2006 International Journal of Oral Biology Vol.31 No.2

Dysplasia-associated seizure disorders are markedly resistant to pharmacological intervention. Relatively little research has been conducted studying the effects of antiepileptic drugs(AEDs) on seizure activity in a rat model of dysplasia. We have used rats exposed to methylazoxymethanol acetate(MAM) in utero, an animal model featuring nodular heterotopia, to investigate the effects of AEDs in the dysplastic brain. Pilocarpine was used to induce acute seizure in MAM-exposed and age-matched vehicle-injucted control animals. Field potential recordings were used to monitor amplitude and numbers population spikes, and paired pulse inhibition in response to stimulation of commissural pathway. Two commonly used AEDs were tested: diazepam 5, 2.5mg/kg;phenytoin 40, 60mg/kg. Diazepam(DZP) and phenytoin(PHT) reduced the amplitude of population spike in control and MAM-exposed rats. However, the amplitude of population spike was nearly eliminated in control rats as compared to the MAM-exposed rats. Pharmaco-resistance was tested by measuring seizure latencies in awake rats after pilocarpine administration(320mg/kg, i.p.) with and without pretreatment with AEDs. Pre-treatment with PHT 60 mg prolonged seizure latency in control rats, but not in MAM-exposed animals. The main findings of this study are that acute seizures initiated in MAM-exposed rats are relatively resistant to standard AEDs assessed in vivo. These data suggest that animal model with cortical dysplasia can be used to screen the effects of potential AEDs.

한국인 갑상선 수질암 환자에서 RET 원종양유전자 점돌연변이 양상

김형훈,김현진,정윤재,민용기,이명식,이문규,김광원,기창석,김종원,정재훈 대한내분비학회 2003 Endocrinology and metabolism Vol.18 No.4

연구배경: 갑상선 수질암의 25∼30%는 유전성으로 발현되는데, MEN 2A, MEN 2B 또는 가족성 수질암의 형태로 나타난다. RET 원종양유전자의 점돌연변이가 유전성 갑상선 수질암의 발생에 중요한 역할을 하므로, 진단 당시의 연령이나 가족력 유무에 관계없이 모든 갑상선 수질암 환자나 또는 RET 변이가 발견된 수질암 환자의 가족 구성원들에서 RET 변이 검색을 하여야 한다. 또한 일부 문헌에서 RET 변이의 양상에따라 임상상이 다르게 표현됨이 보고된 바 있다. 이에 저자들은 한국인 갑상선 수질암 환자에서 RET 원종양유전자의 점돌연변이의 양성률을 알아보고, 변이 양상에 따른 임상상의 차이가 있는지를 알아보고자 본 연구를 시행하였다. 방법: 치근 7년간 본원에서 갑상선절제술을 통해 갑상선 수질암으로 진단받은 29예에서 RET 원종양유전자의 점돌연변이 검사를 시행하였다. 29예의 평균연령은 39세(20∼60세)이었고, 남자 7예, 여자 22예 이었다. 이들의 말초혈액에서 genomic DNA를 분리하고, 특이 시발차를 이용하여 RET 원종양유전자의 exon10, 11, 13, 14, 16부위를 증폭하였다. 증폭된 부위를 자동염기서열분석기를 이용하여 직접 분석하였다. 양성으로 나온 경우는 모든 가족 구성원을 대상으로 RET 변이 유무를 검색하였다. 결과: 대상 환자 29예 중 9예 (31%)에서 RET 원종양유전자 점돌연변이가 발견되었다 RET 변이가 발견된 9예 (남자 3예, 여자 6예)의 평균 연령은 33세 (20∼51세)로 RET 변이가 발견되지 않은 20예의 평균연령 42세(24∼60세)보다 의미 있게 적었다. RET 변이가 발견된 9예 중 MEN 2A가 5예, 가족성 수질암이 1예, 그리고 산발성 수질암이 3예이었고, MEN 2B는단 1예도 진단되지 않았다. MEN 2A 5예 중 4예는 exon 11의 codon 634번(C634R 2예, C634Y 2예)에서, 그리고 나머지 1예는exon 10의 codon 618번 (C618R)에서 변이가 각각 발견되었다. 가족성 수질암 1아는 codon 634번(C634W)에서, 산발성 수질암 3예도 모두 codon 634번 (C634y 2예, C634s 1예)에서 각각 변이가 발견되었다. RET변이 양상 또는 위치에 따른 임상상의 차이는 발견할 수 없었다. 결론: 갑상선 수질암 환자 31%에서 RET 원종양유전자의 점돌연변이를 발견할 수 있었다. 유전성 수질암 중 가족성 수질암 1예를 제외하고 나머지 5예는MEN 2A이었다. 저자들의 5예와 지금까지 국내에서 보고 된 7예를 합친 국내 MEN 2A 12예 중 75% (9/12)는 exon 11의codon 634번(C634R 4예, C634y 4예, C634w 1예)에서, 그리고 나머지 25% (3/12)는 exon 10의 codon618번(C618R 2예, C618s 1예)에서 변이가 발견되었다. 국내에서는 codon 634과 codon 618 두 곳에만 국한된 양상이었고, codon 634에서의 C634R 변이는 1/3에서만 나타났다. 비록 본 연구에서는 제한된 환자 수 때문에 변이 양상과 임상상의 관계를 규명할 수 없었지만, 향후 많은 수의 환자를 대상으로 전향적인 연구를 시행하여 genotype-phenotype 관계 규명을 하는 것이 필요하다. Background: Medullary thyroid carcinomas (MTC) have been reported as hereditary in about 25 -30% of cases. The identification of germline mutation in RET proto-oncogene is important in the diagnosis of hereditary MTC, and occurs in three forms: MEN 2A, MEN 2B and familial MTC (FMTC). To evaluate the prevalence of the relationship of RET proto-oncogene mutation and genotype-phenotype was studied in Korean patients with MTC. Methods: Genomic DNA was obtained from 29 patients, with MTC, who underwent a total thyroidectomy, between 1997 and 2003, at the Samsung Medical Center. There were 7 male and 22 female patients, with an average age of 39, ranging from 20 to 60 years. Exon 10, 11, 13, 14 and 16 of the RET proto-oncogene were amplified, with specific primers, using PCR. A sequencing analysis was performed on the PCR product using an automatic sequencing analyzer. Results: Nine of the 29 patients (31%) were identified as having RET mutations. The average age of these 9 patients was 33 years, ranging from 20 to 51, with a female to male ratio of 2. Five patients had MEN 2A and one had FMTC, with the other 3 thought to have non-hereditary (sporadic) MTC. The 4 patients with MEN 2A had RET mutations on codon 634 of exon 11 (2 patients, C634R; 2 patients, C634Y) and the other patient on codon 618 of exon 10 (C618R). One patient with FMTC had a mutation on codon 634 (C634W). Three patients with sporadic MTC had RET mutations on codon 634 (2 patients, C634Y; 1 patient, C634S). However, no genotype- phenotype relationship could be found, due to the limited number of patients. Conclusion: Thirty-one percent (9/29) of the patients with MTC had RET proto-oncogene mutations. Three-quarters (9/12) of the Korean patients with MEN 2A, including another 7 patients reported in 3 papers in Korea, had RET mutations on codon 634 of exon 11 (4 patients, C634R; 4 patients, C634Y; 1 patient, C634W), but a quarter (3/12) had mutations on codon 618 of exon 10 (2 patients, C618R; 1 patient, C618S). Although no relations could be found between the genotypes and phenotypes, extensive prospective studies will be required to verify this (J Kor SOC Endocrinol 18:360-370, 2003).

생체 인식 정보 검출을 위한 Watershed 기반 Texture 병합에 관한 연구

김진만,김성현,조상현,김태형,김두영 東亞大學校附設情報技術硏究所 2005 情報技術硏究所論文誌 Vol.12 No.2

this paper presents the method that extracts by using watershed algorithm the area which is available for the feature factor in the iris. The inputted image is preprocessed and transferred to the texture block pattern by Polar Coordinate System Transformation. In this paper, the iris feature area is extracted from the image of Texture Block Pattern using Watershed Algorithm. The extracted area is indexed by average value and size of the feature area. After the position information of the feature area is rearranged by Polar Coordinate System Inverse-Transformation. In result, the proposed method showed that is possible to extract the definite iris feature.

Lidocaine 이 흰쥐 해마박편에서 CA 1 추체세포의 활성에 미치는 영향

김형룡,민승기,김대업,오상천,유형근,김은철,고건일,원진숙 원광대학교 생명공학연구소 1995 생명공학연구소보 Vol.3 No.1

Natural Toxin의 안전성 평가연구 : Ochratoxin A의 면역독성 Immunotoxicity of ochratoxin A

김주일,한형미,정혜주,김형수,김환묵,정승태,박재현,강선경,김진호 식품의약품안전청 1997 식품의약품안전청 연보 Vol.1 No.-

Ocllratoxin A는 fenicl'flrium과 rtsr☞rgiffus 속 등의 곰팡of가 생성하는 mycotoxin으로서 세포매기성 및 체액성 면역기능에 슨상을 일으키는 것으로 알려져 있다. 본 연구는 OTA가 면역계에 미치는 영향과 OTA의 면역 독성 억제 효과가 있는 물질을 검색하기 위하며 6~7 주령의 Balb/c mice 비장세핀에 T 및 B 세포의 mitogen인 Con A와 LPS로 처리하고 3일간 배양한 후 비장세포의 분화정도를 MTS를 처리하여 관찰하였으며, OTA를 투여한 마우스의 면역장기 형리조직학적 견화를 관찰하였다. 마우스의 비장세포 (2xtD6cel1/rnf)에 OTA(1~6μg/ml)를 상에서 혼합하여 Con A와 LPS로 자극시킨 결과 임파구는 Sfg/mf이상에서 거의 모든 세포가 분화되지 않았다. mice에 OTA (1,5 및 10mg/11g)를 2일 간격으로 6회 복갚 주사한 후 비장세포를 분리하여 Con A와 LPS로 처리한 결과 Smg 투여군에서는 6회 투여 혼에 분화가 억제되었고, 10mg 투여군에서는 3회 및 6회 투여군에서 분화가 억제되었단. Ochratoxin A의 면역독성 억제 효과가 있는 물질을 검색하기 위하여 in vitro 실험에서 마우스의 비장세포 (2×10" cell/mf)에 OTA (3fg/mf)와 L-phenylalanine, indole-3-carbinol, L-ascorbic acid,piroxicaTn, curcumin 및 arethylsalicylic acid를 혼합하고 동일 처리한 결과 L-ascorbic acid, L-phenylala-nine 및 Indole-3-carbinol은 OTA의 면역독성 발현을 운의성 있게 억제하였다. Balt/c mice에 OTA (10mg/kg)와 l'n pifro실험에서 억제효파가 확인된 L-ascorbic acfd, L-phenylalanine 및 indole-3-carbinol을 2일 간격으로 동시에 3회 복강주사하여 관찰한 결과 indote-3-carbine께서 엮제효과를 관찰할수 있었다. Ochratoxin A를 투여할 마우스의 비장 및 홍선에서는 임파구의 빈도가 줄었으며 특히 비장제서는 starry-sky 세포가 증가됨을 관찰할 수 있었다. 이러한 결자로 OTA는 비장세포의 T 및 B 임파구의 분열능을 억제시 키며, 면역장기의 임파구에 손상 야기시키므로서 면역기능을 저하시키고 OTA에 의한 면역독성은 indole-3-carbinol이 경감시킴을 관찰하엿다. Ochratoxin A (OTA) is a Tnyeotoxin produced by several fungal species iBcluded the genera fs4pergiffug and f☞HiciHiHuL and known as one of the major environmental contaminants. Tn thepresent study, the effects of OTA on the imraune system and the prevention of the OTA-inducedimmunosuppression by protective agents were studied in Balb/c rnicf:. Splenocytes were isolated and theproliferative responses to concanavalin A (Con A) and lipopolysaccharide (LPS) were measured in thepreseEce of 1 to 6fg/mf OTA. Sfg/mf OTA completely btocked both Con A and LPS-stimulatedmitogenic responses, causing approximately 50% inhibition at 3fe/mf OTA. This suppression of mitogen-induced proliferative resplonses observed id uifro was reproduced in mice treated with ☞rA in rioafntraperitoneal administration of 5, IDmg/kg OTA 3 to etimes every other day significantly decreasedboth Con A- aud LPS-inttuced mitegenic responses. The effects of possible protective agents on theOTA-induced suppression c)』 mitogenic responses were examined both in uifro and in viua Amoag 6 pos-sible protective agents (1,-phenylalanine, L-ascorbic acid, indole-3- carbinol, curcumin, firoxicam,acethylsaricylic acid) employed in the present study, L-phenylalanine (100ff), L-ascorbic acid (100#M) and iBdote-3-carbinel t 10Df) showed significantly protective effects on the OTA-iBduced suppres-sion of mitogenic responses id uifro. Howeuer, when the protective of·fects of these agents were examinedin rr'ua only indole-3-carbinol showed significantly protective respoases. It has been ebserved that Iyin-phatic cell population in thc spleen and thymus was decreased in raice given OTA by hematoBylin-eosinstaining. These data indicate that OTA suppresses the cell-mediated and humoral immune functioBsand this OTA-iBduced supflression of immune functions can he alleviated by indole-3-carbinol.

간 세포암에서 VEGF, TGF-β1, b-FGF 발현의 의의

김성용,남충현,주종우,채만규,백무준,이문수,김형철,안현철,김홍수,김창진,김창호 순천향의학연구소 2001 Journal of Soonchunhyang Medical Science Vol.7 No.1

Purpose: Angiogenesis is important for the proliferation and the metastasis of solid tumors. The growth of a solid tumor is widely recognized to depend on the process of neovascularrozation. Without angiogenesis, tumors cease to grow beyond even a few milimeters in diameter. It has been shown that tumor vascular density is an independent prognostic marker in several types of human tumors and is known to correlate with poor prognosis. To date, many angiogenic factors have been identified, such as transforming growth factor-α(TGF-α), transforming growth factor-β(TGF-β), fibroblast growth factor family(FGF), vascular endothelial growth factor(VEGF), platelet derived endothelial cell growth factor(PD-ECGF), tumor necrosis factor-α(TNF-α), and angiogenin. Hepatocellular carcinoma(HCC) is the second most common tumor in Korean males and is known as a typical hypervascular tumor with frequent portal vein invastion. The authors identified the expreesion of VEGF, TGF-β1, and b-FGF in HCC specimens and evaluated the relationship between these growth factors and the clinicopathologic characteristics of HCC. Method: We reviewed the medical records of 30 patients who were diagnosed as hepatocellular carinoma treated with hepatic resection between January 1994 and December 1998 in Soonchunhyang University Chunan Hospital. The selection of the cases was decided according to the condition of paraffin block fixation. The prognostic factors such as age, sex, tumor size, concentration of serum α-fetoprotein, presence of liver cirrhosis, presence of tumor emboli in portal vein, TMN stage, amount of transfusion during the operation, hepatitis B virus(HBV) infection, and Edmonson-Steiner(E-S) grade were investigated. Relationship between the prognostic factors and the immunopathologic expression of the TGF-β1, b-FGF, and VEGF was examined. Result: Thirty patients (24 males, 6 females) were included in the current study. The patient's mean age was 50.6 years and the age ranged from 36 to 65 years. The mean size of the tumor was found to be 5.2cm. All the patients were follewed up for 7 to 63 months. Child's classification A patients were 23(76.7%)cases, B patients were 7(23.3%)cases, and C was none. Immunohistochemical staining of HCC tumor mass in VEGF expression patients were 17(56.7%), b-FGF expression patients were 10(33.3%), and TGF-β1 expression patients were 10(33.3%). VEGF expression or more than one positive expression among the three factors correlated with tumor size and the stage of HCC but did not correlated with other clinicopathological characteristics. TGF-β1 and b-FGF did not correlate with any clinicopathological characteristics. Conclusion: The results suggest that the expression of VEGF or more than one positive expression among the three factors in HCC cells may be a significant prognostic factor of HCC.

Cyclicity Revisited: A Correspondence-Theoreic Approach

Jin-Hyung Kim 한국음운론학회 1999 음성·음운·형태론 연구 Vol.5 No.1

Kim, Jin-Hyung. 1999. Cyclicity Revisited: A Correspondence Theoretic Approach. Studies in Phonetics, Phonology and Morphology 5, 123-146. This paper develops the hypothesis that morphologically-related words are required to be phonologically identical by ranked and violable constraints. In English affixation, pairs of morphologically-related surface forms are linked by output-to-output multiple correspondence relations, and affix-controlled distinct OO₁- and OO₂-identity constraints may force a derived word to deviate from the canonical segmental and stress patterns of English in order to be more like its output base. This approach obviates the traditional derivational analysis that misapplication of phonology in complex words is nothing more than by-product of cyclic rule application. Given multiple correspondence relations, total and partial cyclicity effects are captured in a more direct and unified way within nonprocedural Correspondence Theory. (Korea University of Technology and Education)

요 검사용 스트립의 분광학적 분석을 위한 Computer Simulation

김성철,진병문,김재형,김정배,조진욱,전계록,이승진,김건우 동의대학교 기초과학연구소 1999 基礎科學硏究論文集 Vol.9 No.1

The spectroscopic properties of 9 pads in a urine strip were studied to develop a urine analysis system. From the present work, we determined the wavelength range which could accurately distinguish the degrees of 9 test items. A computer simulation was performed to distinguish the primary color reaction in the urine strips by using the reflectance of the urine strips, the iuminous intensity of light-emitting diodes, and the spectral sensitivity of photodiodes. The simulation results agree well with the experimental results obtained by using a UV-Visible spectrophotometer.

삼중시기 나선형 CT를 이용한 간경변증 환자에서의 결절성 간세포암의 조영증강양상

김영화,윤종필,김형환,이상진,이혜경,김일영,장유진 순천향의학연구소 1996 Journal of Soonchunhyang Medical Science Vol.2 No.2

We evaluated the contrast enhancement pattern of nodular hepatocelluar carcinoma in liver cirrhosis patients by using three phase spiral CT. We anlayzed 34 nodular hepatocelluar carcinonmas in 11 patients with liver cirrhosis. Three phase spiral CT were obtained at 30(arterial phase), 60(portal phase), 180seconds(delayed phase) after initiation of contrast injection in the rate of 3ml/sec. Nodules are classified according to size, as group A(below 1cm, n=11), group B(1 - 3cm, n=15), group C(above 3cm, n=8). Enhancement pattern of the nodule was analyzed comparing to normal liver parenchyme in each phase., as total high, central high, peripheral high, iso, low, mixed density. We also analyzed the detection rate of the nodules. As a whole the nodules show high density in arterial phase(76%), high density in portal phase(47%) and low density in delayed phase(56%). In group A, the nodules show high(100%) in arterial phase, iso(54%) or high (46%) in portal phase, iso(82%) in delayed phase. In group B, the nodules show high(100%) in arterial phase, high(74%) in portal phase, mixed (60%) in delayed phase. In group C, the nodules show mixed density in all three phase. The most common enhancement pattern in each group was high(arterial phase) - iso(portal phase) - low (delayed phase) in group A(6/11, 54%), high - high - low in group B (5/15, 33%), mixed - mixed - mixed in group C(5/8, 63%). The detection rate of the nodule was highest in arterial phase, and the usefulness of arterial phase comparing to portal or delayed phase was higher in smaller size. In conclusion, hepatocelluar carcinomas show different enhancement pattern according to the size on three phase spiral CT, and this may be helpful in diagnosis of them.