고등어에서 분리한 고래회충 유충 : 형태학적 분류, 배양 및 항원성단백질의 분리

안명희,민득영 한양대학교 의과대학 1995 한양의대 학술지 Vol.15 No.2

Eighty six cases of human anisakiasis have been reported since 1971 in Korea, but there might have been more cases of the Anisakis infection due to food habit of this country. This study was conducted to identify morphologic features, changes of larva after cultivation in vitro and antigenic profiles of Anisakid larva and excretory and secretory products. We purchased 27 mackerels from a market in Seoul, dissected the fish and collect the anisakid larvae. Microscopic examination of larvae was done after fixation(FAAG solution) and clearing(lactophenol solution). Total length and width, esophageal length(muscular and ventricular parts), character of head and tail of larvae were observed. Anisakid larvae in PBS solution were froze and thawed, 3∼4 times and the supernatant were collected after centrifugation at 3,000rpm for 30 minutes followed by another centrifugation at 20,000rpm for 1 hour. Larvae were cultured in MEM/5% FCS at 37℃, 5% CO₂incubator. Cultured medium were collected every day and concentrated through Centriplus(Amicon, M. W. ; 10,000) for excretory and secretory proteins. Both antigens were measured protein by Lowry method. Rabbirs were injected with larval antigen mixed with complete adjuvant intramuscularlly, 2 times for 3 weeks interval. Sera were collected before immunization, 3 weeks after 1st immunization, 2, 4, 6, 8, 10, 12 weeks after 2nd immunization. Serum IgG level were evaluated by ELISA method. Anisakid larva and excretory and secretory antigens were identified by immunoblotting. All larvae from the mackerel were identified as Anisakis sp. type I. Anterior end of the worm showed a boring tooth on the lips. Esophagus was formed a muscular part and a ventricular part. Short tail was bluntly ended with a small sized and various shaped mucron. Nerve ring and excretory canal were noted anterior part of the worm. One week after cultivation, many worms were molted and lost boring tooth and tail mucron. Distinctly developed mouth part and rumpled intestine were noted. Cuticular striation was evident compared with that of the larvae before cultivation esp. esophageal region. Two weeks after cultivation, thickened cuticle, sigmoid ventricular part of esophagus, more rumpled intestine were obsered. Activity of the larvae almost stopped on this week. Serum IgG level began to elevate after 1st immunization and prolonged high titer to 12 weeks after 2nd immunization. Protein bands of 199kDa, 68kDa of crude somatic antigens and 100kDa, 77kDa of excretory and secretory(E/S) products reacted with immunized rabbit serum when 7.5% running gel was used on SDS-PAGE. Protein bands of 52 kDa, 20∼19kDa of crude somatic antigen and 50kDa, 45kDa, 23kDa, 23kDa, 20kDa of E/S product reacted with immunized serum on 12.5% running gel. Anisakis larva contained common antigens of Ascaris lumbricoides adult worm.

활성대식세포에 전처리한 톡소포자충의 감염후 증식억제

안명희,민득영 한양대학교 의과대학 1995 한양의대 학술지 Vol.15 No.2

Toxoplasma gondii is an obligate intracellular protozoan parasite. Acute toxoplasmosis in human is a serious infectious disease, especially for pregnant women or immunosuppressed patients. We carried out an experiment to rlucidate the effect of activated macrophage interact with T. gondii (RH) tachyzoites in vitro. Peritonel cells were collected from BALB/c mouse after injection of MEM/10%FCS and incubated with medium in 6 well plate with cover glass at 37℃, 5% CO₂condition for 2 hours. Adherent macrophage cells(1∼5×10 ) were treated with LPS(2.5g/ml, Sigma) or crude lymphokine(immunized mouse spleen cell culture supernatant). Activated macrophages were infected with T. gondii tachzoite which were pretreated with immunized serum, ethyl alcohol, H₂O₂, trypsin, TLCK(tosyl-lysine chlormethyl ketone), triton X-100 and formalin at room temperature for 30 minutes, respectively. Following macrophage and T. gondii interaction (24hrs), cover glass were stained with Giemsa solution. Survival of macrophage and number of intracellular parasite were determined by examining 5 fields of 400X or 1,000X magnification. After pretreatment of T. gondii with ethyl alcohol, H₂O₂, etc., parasite viability were tested by trypan blue dye. Mouse was intraperitoneally injected with pretreated T. gondii tachyzoite(1-5×10 ) and changes of peritoneal cell were observed 1 or 3 days after infection with pretreated T. gondii tachyzoite. Immunized serum treated T. gondii showed supressed macrophage cell rupture and parasite proliferation in cytoplasm. But LPS activated macrophage revealed enhanced cell rupture and tachyzoite multiplication due to enhanced phagocytosis of activated macrophge. T. gondii tachyzoite were damaged after pretreatment of parasite with ethyl alcohol of 50%, H₂O₂of 3.5%, triton X-100 of 0.1%, trypsin of 0.25%, freezing and thawing. Examination of mouse peritoneal cells after i. p. injection of pretreated T. gondii tachyzoites(1-5×10 ) showed no significant changes except live tachyzoite injected control group. As above results, it is considered that infection of macrophage with immune serum treated T. gondii supressed macrophage rupture and intracellular multiplication. T. gondii pretreated with ethyl alcohol, H₂O₂etc. , were damaged and lost the ability to macrophage invasion.

우리나라에서 말라리아의 재출현

안명희,민득영 한양대학교 의과대학 1999 한양의대 학술지 Vol.19 No.1

Malaria is an arthropod bome parasitic disease via Anopheles sp. mosquito. About 200 million malaria cases are reported every year throughout the world. And two million deaths of falciparum malaria per year are serious problems. In Korea, tertian malaria(Plasmodium vivax) has been present with low endemicity until 1940s. During the Korean War, 1950-1953, malaria state in Korea was worsend. After war, National Malaria Eradication Programme was started in collaboration with WHO on 1960. Indigenous malaria was not reported any more in Korea after the mid-1970s except imported cases. One indigenous malaria by Plasmodium vivax was detected in 1993. The patient was a 23-Y-O young man who worked at Military Service in north Kyonggi-do, near DMZ(Demilitarized Zone). The annual number of vivax malaria has increased sharply to 25 in 1994, 107 in 1995, 356 in 1996, 1724 in 1997 and 2632 persons until August 1998. Reemergence of malaria is localized in north Kyonggi-do and Kangwon-do, near DMZ. Patients of 81.4% was army soldiers and veterans. It might be the origin of malaia in South Korea was infected mosquitoes flied from North Korea through DMZ. But we have no information of malaria in North Korea and more precise epidemiologic study will be needed in near future.

질편모충(Trichomonas vaginalis)의 시험관내에서의 용혈능

안명희,류재숙,홍택원,민득영 한양대학교 의과대학 1995 한양의대 학술지 Vol.15 No.2

To evaluate the hemolytic activity of T. vagindlis from Korean women and fresh human RBC of A type were nsed for hemolytic assay. RBC(1×10 /ml) was added to RPMI-TPS media(RPMI 1840:TPS-1= 20 : 1) containing T. vaginalis(2×10 ) and incubated at 37℃, 5% CO₂incubator for 6hr and hemolysis was measured by spectrophotometer at 412m. We investigated the parasite to erythrocyte ratio and lemperature needed for hemotysis. Optimal erythrocyte lysis by T. vaginalis was achieved at ratio of 1 : 5 and optimol temperature 37℃. Minimal hemolytic activity was detected in lysates of T. vaginalis. No hemolysis occured when a membrane a 3㎛ pore size was nsed to prevent contact between parasites and erythrocytes. These data suggest a need for contact for trichomonol hemolysis. Hemolytic activity did not correlated with the production of subcutaneous abscess in mice.

면역시킨 마우스 흉선 및 비장세포에 대한 병원성 Naegleria의 세포병변 효과

안명희,류재숙,민득영 한양대학교 의과대학 1991 한양의대 학술지 Vol.11 No.1

Naegleria fowleri is a causative agent of primary amoebic meningoencephalitis (PAM) in man and experimental animal. We investigated the interaction of immunized mouse thymocyte and splenocyte with N. fowleri trophozoites in vitro. BALb/c mouse was immunized with 150∼200×10⁴/ml N. fowleri trophozoites, intraperitoneally, once a week, for two times. Thymus and spleen cells from immunized or normal mouse cocultured with N.fowleri in 10% FCS+MEM, 37℃, 55%-CO₂incubator and cell: amoeba ratio was 200∼400:1. Thymus or spleen cells and N. fowleri were observed at 24, 48, 72 hrs after incubation by inverted microscope. Spleen cells from immunized mice showed moderate lysis aftre 72 hrs with round amoeba, but other group cells were completely lysed at 48 hrs after incubation with N. fowleri. It may be secreted a certain cytolytic amterial from N. fowleri because of cell lysis occured abruptly after 24 hrs culture. And amoebae showed better condition i.e., pseudopodaand vacuoles, after phagocytosis of lytic cell material. On the other hand, nonadherent cells from immunized spleen were completely lysed at 48 hrs with N. fowleri conculture and immunized thymus cell with aditive activated mactophage showed prolonged cell survival. And so, cooperative reaction of immunized T-lympocyte and macrophage are important to supperss N. fowleri in vitro.

해방 이후 대구·경북 지역 신문연재소설에 대한 발굴 조사 연구 : 격동기(1948-1962) 대구경북지역 신문연재소설을 중심으로

한명환,김일영,남금희,안미영 현대문학이론학회 2004 現代文學理論硏究 Vol.0 No.21

The Aim of this thesis is the research and analysis about the feuilletons of Daegu and Gyeongbug Area in 1945-1963. The study of the Korean modern literature after 1945 should be accompanied with the researches of the local feuilletons that really memorizing the Korean War, and 4.19 & 5 · 16 coup d'etat. We know that feuilletons in 1945-1963 was diverse in lenght and in contents, which contained many different aspects. Not only the dividing of families but also the enlightenment of old-fashioned and new ideology related to democratic & mass psychology was addressed. With theses local feuilletons during 1945-1963 we feel the stories were universe and unique. We ascertain that those feuilletons contain the real codes equal to the real life of average people in this turbulent period of 1945-1963, and those people's anxiety and tension despair through reading newspapers The point of view of 1950's literature needs to be revised to be more comprehensive and affirmative. Through this research and study, we have come to collect memorial raw material of korean literature and culture and to be able to write new korean literary or culture history. Happily we could excavate a lot of raw materials; feuilletons during 1947-1961 unknown to korean academic world-these materials have been divided according to the period ; before war, on war, after war and classified according to subjects in romantic, social, historical novels and traditional chinese stories translated in korean and momorandums and so on. Specially these researches and excavates have acquainted us with facts that most feuilletons in that turbulent period had complicated social meanings and codes. And therefore, we come to know that some memorandum and the notes of reporters in 1949-1961 induced korean people in turbulent periods to experience literature that have been arrived in socialistic reviews of the world Finally, we conclude that a research and excavation on the feuilletons of Daegu & Gyeongbug area during 1945-1961 will have to help understanding the aspect of korean literature in 1950's, by adding to the history of korean literature new texts that will be studied in many ways ' forming the writer's group and the literary circle after 1945, moving traces of writers in war, the present status of tests in that period. But these researches and excavations should be accomplished with researches and excavations the feuilletons of Busan and Gy대ngnam Area in the same period. We also hope that this research and study will spread not only korean peninsula but north korean and overseas.

조제림포킨이 대식세포내 Toxoplasma gondii 증식 억제에 미치는 영향

변순옥,안명희,민득영 한양대학교 의과대학 1993 한양의대 학술지 Vol.13 No.1

The present study was undertaken to elucidate the role of crude-lymphokine or rIFN-r treated macrophage against Toxoplasma gondii in vitro. Peritoneal macrophages were collected from BALB/c mouse treated with10% of proteosepeptone. Normal mouse spleen cells were cultured with con-A(10㎍/ml) for 48hours and the culture supernatant was used as crude lymphokine. For macrophage activation, recombinant IFN-r and crude lymphokine were inoculated before 18 hours of T.gondii infection. After 24 hours of T.gondii infection, the effect of host cell and parasite ration, the cytological changes of macrophages by Giemsa and acridine orange ration, the cytological changes of macrophages by Giemasa and acridine orange stains and NO₂production of activated macrophage were observed. Normal and protenose-peptone treated macrophages were destroyed more severly (76-100%) than those of activated with rIFN-r or crudelymphokine(51-75%). Higher ratio of host cell to parasite revealed the lower destruction rate of macrophgages. Proteose-peptone treated macrophages revealed numerous lysosomal granles in cytoplasm. Activated macrophages produced 7-10 times of amount of NO₂than control by addition of rIFN-r or crude lymphokine and remarkable inhibition of parasite proliferation was observed. The crude lymphokine or IL-2induced slight increase of blastogenesis of immunized mouse spleen cells. With above results, it is assumed that activated macrophages with crude lymphokine produce large amount of NO₂and NO₂was involved in destruction or inhibition T.gondii tachyzoites multiplication in vitro.

간 흡충증에 관한 조사; 한양대학병원에서 진단된 환자를 중심으로

任敬一,安明姬 한양대학교 의과대학 1985 한양의대 학술지 Vol.5 No.1

1983년 한해동안 한양대학병원에 내원한 환자의 총 대변검사 8,561례중에서 187례의 간흡충환자를 찾을 수 있었다. 연령별로는 40대, 50대가 51.3%로써 가장 많았으며 성별비는 5.7 : 1로 남자가 월등히 많았다. 경 또는 중등도 감염이 전체 간흡충증 환자의 87.5%로 대부분이었고 감염정도와 말초혈액의 호산구증다간에는 상관성이 있었다. 입원환자의 주된 동반 질병은 소화기질환이었으며 간흡충증환자에 대한 치료시행이나 치료후 추적연구가 불충분하였다. Total 187 cases of Clonorchiasis were detected out of 8,561 samples which were examined by the formalin-ether concentration method and Stoll's egg counting method, among the patients visiting Hanyang University hospital, from January to December, 1983 in Seoul. This study was undertaken to reveal the change of clinical pattern of Clonorchiasis, comparing with the previous reports. 1. The highest prevalence of Clonorchiasis was found in the 40 - 49 years group. And sex ratio of male vs female was 5.7:1. 2. Light and moderate infection cases, less than 9,999 in E.P.G., were detected 87.1% out of total cases tested. 3. There was a good correlation between the degree of peripheral blood eosinophilia and E.P.G. (coefficient 0.65) 4. Among the associated diseases of inpatients, gastrointestinal diseases were frequently involved. 5. Follow-up study after anthelminthic treatment was carried out unsatisfactorily.

백서 해마에서 카이닌산 유도 경련에 의한 JNK 경유 신호전달경로 활성화의 발달 단계에 따른 변화

김종흔,정희연,노명선,안용민,강웅구,김용식,조수철 大韓神經精神醫學會 2001 신경정신의학 Vol.40 No.5

연구목적 : 어린 백서의 해마에서 전기경련충격(electroconvulsive shock, ECS) 후 활성화되는 것으로 알려져 있는 MAPK계 신호전달경로가 카이닌산(Kainic acid, KA)에 의한 경련 후 어떤 양상으로 활성화되는 지를 초기 발달 단계에 따라서 관찰하고자 하였다. 방 법 : 생후 7일, 14일, 21일 된 어린 백서와 성년 백서의 해마에서 면역블롯 방법으로 MAPK계 효소의 인산화 효소 중 하나인 SAPK/ERK kinase-1(SEK-1), MAPK계 효소인 c-Jun N terminal protein kinase(JNK), 전사인자 c-Jun 및 조기유전자 단백질 Fos의 단백질 양과 기저 인산화를 측정하였고, 백서에게 KA를 주사하여 경련을 유발한 후 위 단백질들의 인산화 변화의 양상을 같은 방법으로 관찰하였다. 결 과 : SEK-1, JNK, c-Jun 모두 연령 증가에 따른 단백질 양 자체에는 변화가 없었다. JNK와 c-Jun은 기저 인산화의 양적 변화도 없었으나 SEK-1의 경우 기저 인산화 양이 생후 7일에 가장 높고 이후 연령이 높아짐에 따라 감소하였다. JNK의 경우 KA 주사 후 인산화 변화 양상이 관찰되지 않았으나 SEK-1과 c-Jun은 생후 14일부터 연령의 증가와 함께 인산화 양도 증가하였다. Fos 역시 생후 7일부터 발현되어 연령의 증가와 함께 발현양이 증가하였다. 결 론 : 백서 해마에서의 MAPK계 신호전달계의 성숙은 연령의 증가에 따라, 또 신호전달 물질에 따라 차이를 보였다. KA 유도 경련 후 JNK는 생후 21일까지도 활성화가 되지 않는 것으로 보아, 세포주에서 알려진 SEK-1 - JNK-c-Jun - Fos로 연결되는 신호전달 경로가 어린 백서의 해마에서는 적용될 수 없음을 시사한다. Objective : We observed the developmental pattern of activation of MAPK signal transduction pathways known to be activated by eletroconvulsive shock(ECS) in young rat hippocampus after kainic acid(KA)-induced seizure. Methods : We used the method of immunoblotting for examining the basal protein amount and basal level of phosphorylation of MAPK kinase(SAPK/ERK kinase -1, SEK-1), MAPK(c-Jun N terminal protein kinase, JNK), transcription factor(c-Jun) and immediate early gene proteins(Fos) in rat hippocampus at postnatal day 7, 14 and 21, respectively. We also examined the changes of phosphorylation of those proteins after kainic acid-induced seizure in the same way. Results : The basal protein amounts of SEK-1, JNK, and c-Jun did not show age-dependent changes and basal level of phosphorylation of JNK and c-Jun remains unchanged throughout the early developmental period. The basal level of phosphorylation of SEK-1 was peaked at postnatal 7 days and then decreased with aging. After kainic acid-induced seizure, the change of phosphorylation of JNK was not observed but those of SEK-1 and c-Jun increased after postnatal day 14. The expression of Fos was observed at postnatal day 7 and also increased with aging. Conclusion : These results show that the MAPK signal transduction system in rat hippocampus matures in accordance with aging, but the process of maturation differs depending specific proteins. This study suggests the signal transduction cascade(SEK-1 - JNK - c-Jun - Fos) which is well established in cell line studies may not be applied to rat hipposcampus because we could not observe the activation of JNK after KA-induced seizure in young rat hippocampus.

영아 돌연사에 대한 고찰 : 13 부검례를 포함한 34례분석 Analysis of 34 Cases Including 13 Autopsies

문여옥,최희경,허정아,신우종,김명아,이성용,장성희,동은실,김종재,안영민,지제근 대한소아과학회 2002 小兒科 Vol.45 No.9