Localisation of DNA Topoisomerase llα in Mouse Erythroleukemia Cells

Ivanova, emilia Ch.,Donev, Rossen M.,Djondjurov, Lalio P. Korean Society of Molecular Biology 1999 Molecules and cells Vol.9 No.3

96년도 상반기에 경상남도 중부지방에서 유행한 무균성 뇌막염에 대한 고찰

권오수,이경림,김원엽,정원조,마상혁,이규만,Kwon, Oh Su,Lee, Kyoung Lim,Kim, Won Youb,Jung, Won Jo,Ma, Sang Hyouk,Lee, Kyu Man 대한소아감염학회 1997 Pediatric Infection and Vaccine Vol.4 No.1

목 적 : 무균성 뇌막염은 소아에서 많이 발병하는 질환으로 여름과 가을에 많이 호발하며 원인은 주로 장바이러스(enterovirus)로 알려져 있다. 감염된 환아들은 단기간에 증상이 좋 아지며 대부분 합병증 없이 회복된다. 이에 저자들은 1996년 5왈말부터 8월까지 마산지역을 중심으로 무균성 뇌막염이 유행하였던 바 뇌막염 환자 및 뇌막염 증상은 보이지 않으나 고열과 발진을 주소로 내원한 환아에서 원인 바이러스를 규명하고 임상 증상과의 관계를 추구 하고자 본 연구를 시행하였다. 방 법 : 1996년 5월부터 8월까지 마산파티마 병원에 무균성 뇌막염으로 진단된 환아 및 고열이 나면서 몸에 특징적인 발진이 생겨 바이러스 감염이라고 생각되었던 환아 155명중 57명에서 검체를 채취하여 (뇌척수액22례, 대변57례) 바이러스 배양과 장바이러스 (enterovirus) RNA의 검출을 시행하였다. 뇌척수액은 요추천자를 하여 무균 튜브에 채취하였고 대변은 면봉법으로 채취하여 무균 튜브에 넣어서 검사 전까지 $-30^{\circ}C$ 냉동고에 보관하여 검사실까지 운반시는 밀봉된 용기에 드라이아이스를 넣어 이송하였다. 채취된 뇌척수액과 대변에서 바이러스 분리를 시행하고 면역형광법을 이용하여 바이러스를 동정하였다. 바이러스 배양에 사용한 세포주들은 human embryonal rhabdomyosarcoma(RD), African green monkey kidney(BS-C-1), human lung diploid(MRC-5), human cervix epidermoid carcinoma(HeLa)세포주였고 뇌척수액에서의 장바이러스 RNA는 reverse transcription-polymerase chain reaction(RT-PCR)을 이용하여 확인하였다. 결 과 : 1) 바이러스 감염으로 인하여 입원하였던 환아는 155명이었으며 남녀의 비는 1.94:1로 남아가 많았고 연령분포는 생후 15일부터 15세까지 다양한 연령층의 분포를 보였으나 주로 5세미만에서 발생하였으며(74.8%) 시기상으로는 1996년 5월부터 1996년 8월까지 환자가 발생 하였다. 2) 임상 증상은 전례에서 고열이 동반되었으며 그 외에 두통, 구토, 복통, 설사, 발진이 주된 임상 증상이었으며 발진은 4세 미만에서, 특히 18개월 이하에서 많이 관찰되었다 (p<0.001). 3) 뇌척수액을 이용한 RT-PCR 결과는 22례 중 10례에서 양성의 결과가 나왔고 그중 3례에서 echovirus type 9이 분리되었다. 4) 대변검체가 의뢰된 환자 54례중 41례에서 바이러스가 분리되었으며 분리된 바이러스는 모두 echovirus type 9 이었다. 결 론 : 1996년도 상반기에 경상남도 중부 지방에서 유행한 무균성 뇌막염의 원인 바이러스와 같은 시기에 유행한 고열과 발진의 원인 바이러스는 동일한 바이러스로서 모두 echovirus type 9 이었다. Purpose : Aseptic meningitis mainly caused by enterovirus is common in pediatric population especially during summer & fall. Most of pediatric patients restore their health without any complications with proper management. Between May to August of 1996, Masan and surrounding areas of the Kyoungsangnamdo were epidemic areas for the aseptic meningitis. The purpose of this study was to determine causative virus and describe correlation between disease and clinical symptoms in aseptic meningitis patients and those with fever and characteristic rashes without apparent meningitis symptoms. Methods : Between May to August, 1996, 57 patients with high fever and characteristic feature of rashes were reviewed. From 22 cerebrospinal fluid & 57 stool obtained specimens, viral culture and detection of enterovirus RNA were conducted. Collected specimens were kept in $-30^{\circ}C$ environment until sending of specimens to labortory. The virus identified through indirect immunofluorescence. RT-PCR method was used to identify enterovirus RNA in cerebralspinal fluid. Results : 1) One hundred fifty five pediatric patients with viral infection required hospitalization. Disease occurred higher rate in male than female with ratio of 1.94:1. Examined patients' age ranged from 15days old to 15years old. But most of patients(74.8%) were under age of 5years old. The time of occurrence was between May to August of 1996. 2) All patients had high fever and physical symptoms in those patients include headache, vomiting, abdominal pain, diarrhea, and rashes. The rashes observed mainly in patients under age of 4 years and were predominantly commom patients under age of 18 months olds)<0.001). 3) Between sampled patients and non-sampled patients, clinical course was similar. Echovirus type 9 was cultivated in 41 out of 57 cases of collected stool specimens. RT-PCR that used on CSF showed positive results in 10 out of 22 cases. Three cases of positive cultivated of positive results in RT-PCR were echovirus type 9. Conclusions : Echovirus type 9 was thought to be the causative agent of aseptic meningitis that was prevalent throughout mid areas of Kyoungsangnamdo from May to August, 1996. Additionally causative agent that responsible for high fever with rashes without meningitis symptoms also thought to be the same echovirus type 9.

심상성 천포창 환자에서 자가항체 검출을 위한 간접면역형광법에 기질이 미치는 영향

이상진(Sang Chin Lee),이준영(Jun Young Lee),김영환(Yung Hwan Kim),김정원(Chung Won Kim) 대한피부과학회 1993 大韓皮膚科學會誌 Vol.31 No.4

Background : Substrates play an important role in the effectiveness of the indirect immunofluro-rescence test. The monkey eaophagus has been known to be a more reliable and reproducible substrate than any other animal substrates for detecting pemphigus vulgaris autoantibodies, but it is quite difficult to get the monkey esophagus in Korea, therefore, we tried to find other substrates which could be substituted for it. Objective : We studied to select the substrate which can be the most sensitive, to compare the immunofluorescent sensitivity of various animal substrates and fores,cins for detection of pemphigus autoantibodies, and finally to evaluate the effectiveness of cultured hunan keratinocytes as a substrate for indirect immunofluorescerice. Method : With foreskin, guinea pigs, cats, rats, dogs, rabbits and cultured human keratinocytes as substrates, an indirect imrnunofluorescence test was performed to detect pemphigus vulgaris autoantibodies in seven sera from the active patients before treatment and diluted serially from 1:10 to 1:1280 Result : We could detect pernphigus autoantibodies in all animal subtrates used in this study except rats and obtain good immunofluorescent results with foreskin, guinea pigs, and cultured human keratinocytes. The results were moderate with dogs, cats, rabbits, and poor with rats. Conclusion : We confirm thut cultured human keratinocytes can be a good effective substrate. According to effectiveness, subtrates can be divided into three groups. Foreskin, guinea pigs, and cultured human keratinocytes are included in the good effective substrate group, dogs, cats, and rabbits in the modeate group and rats in the poor substrate group, but we can not find any superiority of substrates within each groups. (Kor J Dermatol 1993;31(4):7474-480)

표피 - 진피 분리 방법에 따른 자가 면역 수포성 질환의 항원성

이범주(Beom Joo Lee),김수찬(Soo Chan Kim),안성구(Sung Ku Ahn),이승헌(Seung Hun Lee) 대한피부과학회 1993 대한피부과학회지 Vol.31 No.1

There are many known methods of dermo-epidermal separation for the investigation of autoimmune blistering diseases. Investigators should select a proper method since many differences exist preservation of antigenicity. In order to determine the stabilization of antigenirity by different separation methods, we have separated dermo-epidermal junction by means of 1M s;ilt, 56C PBS, 20mM EDTA and dispase. Indirect immunofluarescence and immunoblotting were performed on each specimen with sera of patients with pemphigus vulgaris, pemphigus foliaceus, paraneoplastic pemphigus, bullous pemphigoid and epidermolysis bullosa acquisita. The results are as follows : 1. In indirect immunofluorescence study of pemphigus group, best, result were obtained when normal skin without dermo-epidermal separation was used. Dispase well preserved antigenicity of pemphigus after dermo-epidermal separation, but no differences were noted in antigenicity stabilization among separation mehods by immunoblotting. 2. In indirect immunofluorecence study for differentiation of bullous pemphigoid and epidermolysis bullosa acquisita, we recommend EDTA and dispase methods in addition to 1M salt induced skin separation that have been most popularly used. 3. Results of the immunoblotting of bullous pemphigoid showed that 1M salt, EDTA and heat preserved the antigenicity well but the antigenicity was lost by dispase. 4. Results of the immunoblotting of epidermolysis bullosa acquisita she wed that antigen did not exist in epidermal extract. 5. Antigen preservation according to the different methods of demo-epidermal separation was not identical between indirect immunofluorescence and immunoblotting (Kor J Dermatol 1993; 31 (1): 19-27)

간접 면역형광 검사와 면역 전사 검사를 이용한 천포창 자가항체 검출방법의 감수성 비교

정성재 ( Seong Jai Jeong ),이창우 ( Chang Woo Lee ) 대한피부과학회 1996 大韓皮膚科學會誌 Vol.34 No.2

Background: Immunolcgical assays are required for the accurate diagnosis of autoimmune bullous dermatoses including pemphigus vulgaris(PV) and pemphigus foliaceus(PF). In the detection of circulating autoantibodies to pemphigus antigens(desmosomal components), the priority remains controversial between indirect immunofluorescence(IF) and immunoblot(IB) assay. Objective : In the present study we compared the sensitivity of indirect IF and that of IB using amplified alkaline phosphatase staining system in the detection of pemphigus autoantibodies. Patients : We selected eight patients with serum endpoint titer of 1:80 in preliminary study. Among these patients three were PV and five were PF. Methods/Results : The titers of IgG autoantibodies found on indirect IF were confirmed as 1: 80 in all patients, whereas the titers examined by IB assay were much higher, 1: 640 to 1: 2560. In the 3 sera of PV patients, the titers of two cases were 1: 1280 and the third case was 1: 2560. In 5 cases of PF, one was 1:640, two were 1: 1280, and two were 1:2560. Conclusion : This result suggests that the immunoblot examination using amplified alkaline phosphatase staining system demonstrates higher sensitivity compared with indirect IF(p=0.0003 by Mann-Whitney U test) in the detection of pemphigus autoantibodies. (Kor J Dermatol 1996;34(2): 212-218)

간접면역형광법을 이용한 항핵항체검사에서 Dense Fine Speckled 양상의 임상적 의의

강소영,이우인 대한진단검사의학회 2009 Annals of Laboratory Medicine Vol.29 No.2

Background : Dense fine speckled (DFS) pattern in antinuclear antibody (ANA) test using indirect immunofluorescence method became to be known recently and it is detected in patients with various chronic inflammatory diseases as well as in healthy individuals. We investigated the relation between DFS pattern and various diseases. Methods : ANA tests by indirect immunofluorescence method using HEp-2 cell line slide (Kallestad; Bio-Rad, USA) were performed in 2,654 patients for screening of systemic autoimmune diseases. The frequencies of ANA and DFS positivity were analyzed according to sex, age, clinical department and disease. Results : ANA was positive in 13.3% (352/2,654) of the total patients, and the frequency of DFS pattern was observed in 3.8% (101/2,654) of the total patients and in 28.7% (101/352) of the patients with ANA positivity. Higher frequency of DFS positivity was observed in patients referred from Departments of Rheumatology and Nephrology, but there was no difference in the frequencies of DFS positivity among the patients with ANA positivity. The frequency of DFS pattern was higher in seborrheic dermatitis (14.3%), herpes zoster (11.1%), rheumatoid arthritis (16.9%), systemic lupus erythematosus (15.4%) and Sjogren syndrome (14.3%). Conclusions : The DFS pattern is a frequent finding (about 28% of ANA positivity) in ANA test using indirect immunofluorescence method. Relatively high frequency of DFS pattern was observed in autoimmune diseases, contrary to the previous observations that DFS pattern is not related with autoimmune diseases. Further studies including the confirmation tests of anti-DFS70 are needed for the identification of relation between DFS pattern and particular diseases. 배경 : 최근 간접면역형광법을 이용한 항핵항체(ANA) 검사에 서 특징적인 dense fine speckled (DFS) 양상이 각종 만성염증 성질환 외에도 정상인에서까지 발견되는 것으로 알려졌다. 저자 들은 ANA 검사가 의뢰된 환자들에서 DFS 양상을 보이는 환자 들의 빈도를 조사하고, 임상과별 및 질환별로 DFS 양상을 보인 환자들을 분석하여 특정 질환과의 연관성이 있는지 살펴보고자 하였다. 방법 : 전신자가면역질환의 선별검사 목적으로 ANA 검사가 의뢰된 2,654명의 환자를 대상으로 HEp-2 세포 슬라이드(Kallestad TM; Bio-Rad, Redmond, WA, USA)를 사용한 간접면역 형광법으로 ANA 검사를 시행하였다. ANA 양성 및 DFS 양성 빈도를 성별, 연령별, 임상과별 및 질환별로 분석하였다. 결과 : 2,654명 중 DFS 양성은 101명(3.8%)으로 나타났고 이 는 ANA 양성(352명, 13.3%)의 28.7%인 것으로 나타났다. 임상 과 간 비교에서 DFS 양성 빈도는 류마티스내과 및 신장내과에 서 높았고, ANA 양성자 중 DFS 양성 빈도의 차이는 관찰되지 않았다. 피부과의 경우 지루피부염(14.3%) 및 대상포진(11.1%) 환자에서 DFS 양성 빈도가 높았고, 류마티스내과의 경우 류마 티스관절염(16.9%), 전신홍반루푸스(15.4%) 및 쇼그렌증후군 (14.3%)에서 DFS 양성 빈도가 높았다. 결론 : DFS 양상은 간접면역형광법을 이용한 ANA 검사에서 양성 건수의 약 28%를 차지할 정도로 빈번히 관찰되는 소견으 로 나타났다. DFS 양상과 자가면역질환과의 연관성이 없다는 기존의 보고와 달리 본 연구에서 높은 양성률을 보이므로 특정 질환과의 연관성 유무를 확인하기 anti-DFS70 확인검사를 포 함하는 연구가 뒤따라야 할 것으로 판단된다.

치근단 병변에서 Axtinimyces종의 검정을 위한 간접 면역형과법적 연구

윤수한,장원정,권오양 大韓齒科保存學會 1996 Restorative Dentistry & Endodontics Vol.21 No.1

증식성 천포창 (天疱瘡) 의

김성훈(Sung Hun Kim),원영호(Young Ho Woon),김영표(Young Pio Kim) 대한피부과학회 1988 대한피부과학회지 Vol.26 No.3

We experienced a 54-year-old female patient who had multiple vegetaing plaques and, peripherally, a few pustules in her both axillae. On the skin biopsy specimen of the vegetating plaque, it showed acanthosis, suprabasal acsntholysis, papillo-matosis, downward proliferation of the epidermis, and eosinophilic microroabscess in the epidermis. The indirect immunofluorescence study disclosed anto-antibody positive to intercellular substance of the lip of guinea pig, while direct. immuno-fluorescence of the perilesional skin reealed no specific findings. The skin lesion had improved with topical and systemic steroid therapy.

Performance of an Automated Fluorescence Antinuclear Antibody Image Analyzer

유인영,오종원,차훈석,고은미,강은숙 대한진단검사의학회 2017 Annals of Laboratory Medicine Vol.37 No.3

Background: The gold standard for antinuclear antibody (ANA) screening is the indirect immunofluorescence (IIF) assay with human epithelial cells (HEp-2). However, a number of substantial disadvantages of manual IIF assays have highlighted the need for the automation and standardization of fluorescent ANA (FANA) testing. We evaluated the performance of EUROPattern Suite (Euroimmun AG, Germany), an automated FANA image analyzer, with regard to ANA detection and pattern recognition compared with conventional manual interpretation using the fluorescence microscopic IIF assay. Methods: A total of 104 samples including 70 ANA-positive sera and 34 ANA-negative sera collected from September to October 2015 were included. The sensitivity, specificity, and pattern recognition function were evaluated to determine the performance of EUROPattern Suite compared with the manual IIF assay results. Results: The sensitivity and specificity of EUROPattern Suite for ANA detection were 94.3% and 94.1%, respectively. The concordance rate between the two methods was 94.2%. For pattern recognition, 45.7% of the samples were assigned identical ANA patterns including simple and mixed. When major pattern matching was considered, 83.7% (41/49) and 95.2% (20/21) of the samples with simple and mixed patterns, respectively, showed concordant results between the two methods. Conclusions: EUROPattern Suite, an automated FANA image analyzer, provides a viable option for distinguishing between positive and negative results, although the ability to assign specific patterns is insufficient to replace manual microscopic interpretation. This automated system may increase efficiency in laboratories, in which a large number of samples need to be processed.

쯔쯔가무시병의 진단에 사용되는 피동혈구응집법과 간접면역형광법의 비교 연구

이창섭,권근상,나상언,이흥범 대한감염학회 2008 Infection and Chemotherapy Vol.40 No.1

The gold standard for the serologic detection of scrub typhus is the indirect immunofluorescence assay (IFA), however the passive hemagglutination assay (PHA) is commonly used method in Korea because of its availability, easy handling. We compared the sensitivity of PHA and IFA to compare their effectiveness in diagnosing scrub typhus. Out of 35 patients who were eventually diagnosed of scrub typhus by positive IFA, 19 patients (54%) were also tested positive for PHA during the acute phase (titer=1:80). Whereas 30 patients (85%) were positive in IFA during the acute phase. During the recovery phase 30 (85%) patients were positive in PHA, whereas all patients were found to be positive in IFA. Patients with 4-fold or greater rise of antibody were 12 (34.3%) in PHA, and 19 (54.3%) in IFA. We note that IFA has better sensitivity over PHA in detecting scrub typhus.