Differential Manifestation of Ocular Phenotypes in TALEN-mediated p19arf Knockout FVB/N and C57BL/6J mouse lines

Jin‑Sung Park,Joo‑Il Kim,Hyun‑Jin Lim,Soo‑Kyung Ryu,Euna Kwon,Kang‑Min Han,Ki‑Taek Nam,Han‑Woong Lee,Byeong‑Cheol Kang 한국유전학회 2020 Genes & Genomics Vol.42 No.9

Background p19arf, primarily known as a tumor suppressor, has also been reported to play an essential role in normal development of mouse eyes. Consistently, lack of p19arf has been associated with ocular defects, but the mixed background of the knockout (KO) mouse strain used raised a concern on the accuracy of the phenotypes observed in association with the targeted gene due to genetic heterogeneity. Object We carried out a study to investigate into the efect of genetic background on the manifestation of p19arf KO associated phenotypes. Methods We characterized the phenotypes of novel p19arf KO mouse lines generated in FVB/N and C57BL/6J using a transcription activator-like efector nuclease (TALEN) system in comparison to the reported phenotypes of three other p19arf-defcient mouse lines generated using homologous recombination. Results Ninety-fve percent of FVB/N-p19arf KO mice showed ocular opacity from week 4 after birth which worsened rapidly until week 6, while such abnormality was absent in C57BL/6J-p19arf KO mice up to the age of 26 weeks. Histopathological analysis revealed retrolental masses and dysplasia in the retinal layer in FVB/N-p19arf KO mice from week 4. Besides these, both strains developed normally from birth to week 26 without increased tumorigenesis except for a subcutaneous tumor found in a C57BL/6J-p19arf KO mouse. Conclusion Our fndings demonstrated surprisingly variable manifestation of p19arf-linked phenotypes between FVB/N and C57BL/6J mice, and furthermore between our mouse lines and the established lines, indicating a critical impact of genetic background on functional study of genes using gene targeting strategies in mice.

정상과 갑상선 종양조직에서 사람 IGF-I 유전자의 발현

김성운,장현하,박상미,김덕윤,우정택,양인명,김진우,김영설,김광원,고석환,홍성화,최영길 경희대학교 유전공학연구소 1993 遺傳工學論文集 Vol.5 No.-

Many of the growth-promoting properties of growth hormone(GH) are mediated by insulin-like growth factor-I(IGF-I), a highly conserved circulating 70-amino acid peptide. Recent studies have shown that multiple mechanisms influence IGF-I gene expression, including transcription from two promoters, alternative RNA splicing, and variable polyadenylation. In thyroid tissue, thyroid stimulating hormone(TSH) and IGF-I are the most possible candidates for follicular cell proliferation and hypertrophy. Actually IGF-I had autocrine and paracrine effect for tissue growing. We prepared thyroid tumor tissue mRNAs using single step method for detecting IGF-I levels according to different tissues, i.e., thyroid adenoma or papillary thyroid carcinoma. We used Northern blot analysis for IGF-I mRNA and RNase protection assay (RPA) for IGF-I transcription start sites. For Northern blot, we used whole human IGF-I cDNA as a DNA probe and for RPA, we used IGF-I exon 1 containing noncoding promoter 1 as a riboprobe. We got good RNA bands from Northern blot analysis around 1 kb (IGF-IA) and 7.5 kb (IGF-IB) region. To clarify the amount of both IGF-IA and IB mRNAs, we measured autoradiographied signal of IGF-I mRNAs bands using densitometer. In IGF-IA signals, there's no change among liver and thyroid tissues, but in case of IGF-IB mRNA bands, the signal was markedly increased in thyroid carcinoma tissues than that of normal thyroid tissue (85% vs 14%). In the study of RPA, all thyroid tissues used the same transcription start sites as those of liver's. We concluded that that this different regulation of IGF-I mRNA was originated from tissue specificity. That meant some tissue specific transcription factor/s were related to tissue IGF-I expression.

인슐린 주사제의 고충실 액체색층기 분석

우정택,팽정렬,김성운,양인명,김진우,김영설,김광원,최영길 최신의학사 1993 最新醫學 Vol.36 No.8

세포질내 정자주입법(ICSI)에 있어서 정자흡입 및 난자내 주입방법에 관한 연구

이택후,김항진,송건호,김대근,전상식,박윤규,서태광,전병균,류은경,이은숙,문진수,김광철 경북대학교 의학연구소 2000 경북대학교병원의학연구소논문집 Vol.4 No.1

Study on Method of Sperm Aspiration and Injection into an Oocyte in Intracytoplasmic Sperm Injection(ICSI) Immobilization of spermatozoa prior to intracytoplasmic sperm iniection(ICSI) sometimes results in crooked tail and this makes it difficult to aspirate sperm into an injection pipette tail first. Head-first sperm aspiration into an injection pipette avoid this problem due to the bigger size of the sperm head. The effect of head or tail-first sperm injection into an oocyte on fertilization cleavage, percentage of grade I embryos and development to blastocyst stage in ICSI program has been studied. A single living immobilized spermatozoa from oligoasthenozoospermic patient was injected into an oocyte head-first or tail-first according to the treatment. Eighteen hours after microinjection, oocytes ware inspected for survival and fertilization Fertilized oocytes with two pronuclei were cultured in 30μl drop of mHTF supplemented with 10% heat-inactivated follicular fluid(FF) at 37℃. On day 2. embryo transfer was performed with cleaved embryos. The remaining 2-8 cell stage embryos were co-cultured with BRL cells in mHTF + 10% FF for 72 hours and the developmental stage was observed. The data were analyzed by Analysis of Variance. A total of 164 oocytes from 36 cycles were assigned to earth treatment and ICSI was performed(88 head-first, tail-first). The rates of normal fertilization were 81.8% and 76.3% for head-first and tail-first, respectively. Of the fertilized oocytes, the percentage of cleaved embryos and the percentage of grade 1 embryo among cleaved embryos were 88.9% and 68.8%, 93.1% and 74.1% for head-first and tail-first, respectively. Of the 2-8 cell embryos cultured, 44.4%(16/36) and 50.0%(10/20) for head first and tail first, respectively developed to blastocyst stage. There were no differences in fertilization, cleavage, rates of grade 1 embryos, and development to blastocyst stage. In conclusion, head-first or tail-first sperm injection into an oocyte in ICSI program does not affect fertilization and subsequent embryo development to blastocyst stage in vitro.

심한 골병변으로 발현된 기능성 낭종성 부갑상선 선종

전숙,김영희,박지영,고관표,박철영,김덕윤,우정택,김성운,김진우,김영설,고석환 대한내분비학회 2003 Endocrinology and metabolism Vol.18 No.2

낭종성 부갑상선 선종과 심한 골병변을 동반한 부갑상선 기능항진증은 매우 드문 질환으로서, 저자들은 양측 고관절의 통증을 초기 주소로 내원한 환자에서 고칼슘혈증과 부갑상선 호르몬 증가, 골병변의 방사선적 소견을 통해 부갑상선 기능항진증을 진단하고, 경부 초음파와 컴퓨터 단층 촬영, 부갑상선 스캔검사 및 수술중 부갑상선 낭종액 검사 등을 통해 기능성 부갑상선 낭종의 한 종류인 낭종성 부갑상선 선종을 진단하고 수술적 제거를 통하여 정상화된 1예를 경험하였다. A cystic parathyroid adenoma is rare. A case of primary hyperparathyroidism, with the cystic formation of a parathyroid adenoma and a severe bony lesion, is reported. A 52-year-old male was admitted due to pain in both hips and for evaluation of hypercalcemia. The plasma level of the intact parathyroid hormone (iPTH) was elevated to 1424 pg/mL. Ultrasonography and the computed tomography revealed a parathyroid cyst on the left thyroid lower pole. Parathyroid scintigraphy detected a parathyroid adenoma. A radiograph showed a subperiosteal bone resorption on the phalanges, and a brown tumor (osteitis fibrosa cystica) on the femur shaft was noted. A surgical excision of the parathyroid adenoma was performed. The PTH level in the cystic fluid was increased. A histological examination confirmed a cystic parathyroid adenoma. The PTH level was normalized after the operation (J Kor SOC Endocrinol 18:214-220, 2003).

제2형 당뇨병 남성 환자에서 유리 테스토스테론과 성호르몬 결합 글로블린 농도

남기덕,김영설,박철영,오승준,김덕윤,우정택,김성운,양인명,김진우,최영길 대한당뇨병학회 2002 Diabetes and Metabolism Journal Vol.24 No.6

연구배경:인슐린저항성은 제2형 당뇨병과 심혈관질환의 주요한 위험 인자로 성호르몬과 상호 관계가 있다고 알려져 있다. 그러나 여성과는 달리 남성에서는 연령과 인슐린저항성에 따른 유리 테스토스테론과 성호르몬 결합 글로블린 농도의 변화에 대한 연구가 부족한 실정이다. 그러므로, 본 저자등은 제2형 당뇨병 남성 환자에서 정산인과 비교해서 유리 테스토스테론과 성호르몬 결합 글로블린 농도를 측정하고 연령에 따른 변화 정도를 알아보고자 하였다. 방법:대상 환자 모두에서 연령과 체질량지수, 총 콜레스테롤, 중성지방, 공복혈당과 인슐린 농도를 측정하였다. 혈중 유리 테스토스테론 농도는 방사면역측정법(radioimmunoassay)을 이용해서 측정하였고, 혈중 성호르몬 결합글로불린은 면역방사계측측정법(immunoradiometric assay)을 이용해서 측정하였다(Diagnostic System Laoratories, Wbster, TX, USA). 결과:1)제2형 당뇨병 남성 환자에서 정상 대조군 남성에 비해 성호르몬 결합글로불린은 104.1±35.0 vs 25.7±3.5 mole×10??로 의미 있게 높았으나(p<0.001), 유리 테스토스테론은 13.7±9.5 vs 13.6±6.5 ng/dL로 차이가 없었다. 2)연령과 성호르몬 결합 글로블린 사이의 상관 계수는 제2형 당뇨병 남성 환자에서 0.40로 중등고의 양의 상관 관계를 보였고(p<0.001), 정상 대조군 남성에서 0.11로 유의한 상관 관계를 보이지 않았다. 연령과 유리 테스토스테론 사이의 상관 계수는 제2형 당뇨병 남성 환자에서 0.08, 정상 대조군 남성에서-0.17로 모두에서 유의한 상관 관계를 보이지 않았다. 3)연령과 체질량지수를 보정한 후에 제2형 당뇨병 남성 환자와 정상 대조군 남성에서 혈중 인슐린 농도, 유리 테스토스테론과 성호르몬 결합 글로블린 사이에는 상관 관계가 없었다. 결론:제2형 당뇨병 남성 환자에서 정상 남성에 비해 성호르몬 결합 글로블린 농도가 증가되어 있었으며, 유리 테스토스테론은 차이가 없었다. 연령이 증가함에 따라 성호르몬 결합 글로블린이 제2형 당뇨병 남성 환자에서 정상 남성에 비해 증가 폭이 의미있게 컸으며, 유리 테스토스테론은 변화가 없었다. Background: Insulin resistance is a risk factor for cardiovascular disease and type 2 diabetes mellitus. There are many previous studies indicating that insulin lowers serum sex hormone-binding globulin levels, and there is inverse correlation between insulin resistance and serum sex hormone-binding globulin levels in women. However, in men, a limited number of studies are available to explain the effect of sex hormone on age and insulin. Therefore, the present study was undertaken to investigate the relationship among free testosterone, hormone-binding globulin and age in type 2 diabetic men and control subjects. Method: Age, body mass index, total cholesterol, triglyceride, fasting blood sugar, and insulin concentrations were examined on 89 type 2 diabetic men and 47 control subjects. The free testosterone level was measured by commercially available double-antibody system (Radioimmunoassay). The sex hormone-binding globulin level was also measured by commercially available double-antibody system(Immunoradiometric assay). Results: 1) Sex hormone-binding globulin level was significantly increased in patients with type 2 diabetes. However, there was no significantly difference in free testosterone level between the two groups. 2) Sex hormone-binding globulin was positively correlated with age (r=0.4, p <0.001) in patients with type 2 diabetes. Sex hormone-binding globulin and free testosterone were not correlated with age in control sujects. 3) Free testosterone and sex hormone-binding globulin concentrations were not significantly related to serum insulin concentration after adjusting for age and body mass index. Conclusions: We observed increased sex hormone-binding globulin concentration in diabetes man, and was a positively related to age. Further studies are needed to understand the relationships between age, insulin resistance, testosterone, and sex hormone-binding globulin concentrations(J Kor Diabetes Asso 24:699~707,2000).

당뇨병 백서의 간세포에서 Glucokinase 활성도 및 유전자 발현에 대한 인슐린의 영향

강성이,팽정령,서광식,안규정,우정택,김성운,양인명,김진우,김영설,김광원,최영길 경희대학교 유전공학연구소 1993 遺傳工學論文集 Vol.5 No.-

목적 당대사의 조절 상태에 따른 생체 변화를 분자 수준에서 이해하고자 식이 조건을 달리한 정상 백서와 화학적으로 유도된 당뇨병 백서의 간조직에서 혈당수준과 인슐린치료 정도에 따라 나타나는 글루코키나제 활성도 및 유전자 발현을 분석하였다. 방법 스트렙토조토신 정맥투여 후 당뇨병의 유발을 확인하고, 인슐린을 1일 3회 3일간 복강내로 투여하여 상태를 안정시킨 후, 인슐린 투여군은 인슐린 투여 6시간이내에 그리고 인슐린 비투여군은 24시간 후 단두하여 채혈하고 복강을 열어 간조직을 채취하였다. 채취한 간조직에서 글루코키나제 활성도는 인산화된 포도당에서 NADH의 형성을 형광분광계로 측정하였으며, 글루코키나제 유전자 mRNA발현은 Northern 분석법을 이용하였다. 성적 정상 백서에서 공복상태와 식이를 섭취한 경우에 간조직의 글루코키나제 효소의 활성은 차이가 없었으나, 글루코키나제 유전자 mRNA 발현은 증가되었다. 당뇨병이 유발된 백서의 간조직에서 글루코키나제 효소의 활성 및 글루코키나제 유전자의 mRNA 발현은 정상 백서에 비하여 낮았다. 인슐린 투쳐 후 글루코키나제 효소의 활성 및 글루코키나제 유전자의 mRNA 발현이 증가되었고, 특히 혈당이 정상화된 경우에서 글루코키나제 유전자의 mRNA 발현이 증가도었다. 결론 인슐린에 의한 간조직에서 글루코키나제 효소의 활성 및 글루코키나제 유전자의 mRNA을 증가를 볼수 있었다. 당뇨병 백서에서 인슐린 투여 후에 혈당조절이 안된 경우 간조직의 글루코키나제 유전자의 mRNA 발현이 증가가 없는 것으로 보아 글루코키나제 mRNA의 발현에는 인슐린 이외의 다른 요소가 관여할 것으로 생각된다. The liver-specific hexokinase isoenzyme, referred to as glucokinase, is thought to play a key reglulatory role in hepatic glucose metabolism. The glucokinase gene is, therefore, of interest both because of its tissue-specific expression and because of the several regulatory processes that can be analyzed. The level of hepatic glucokinase activity appears to be determined essentially by regulation of the rate of enzyme synthesis, with insulin playing a leading role as an inducer. We investigated the role of insulin for the induction of glucokinase in the liver of diabetic rats. Experimental diabetes was induced by injection of streptozotocin 7 days before the experiment. Regular insulin was given by three days intraperitoneal injection at 8-h interval. The glucokinase mRNA in the liver was estimated by Nothern blot assay, as well as by fluorometric enzyme activity assay. Glucokinase activity was not reduced in the liver of normal fasting rats as compared to normal fed rats. And glucokinase activity was reduced in the liver of diabetic rats as compared to normal rats. In diabetic rats treated with insulin, glucokinase enzyme activity were increased. But glucokinase mRNA expression was only increased in normoglycemic diabetic rat with treated with insulin as compared to hyperglycemic rat. These data indicate that insulin stimulates hepatic glucokinase enzyme activity and mRNA expression. But other hormonal or metabolic factors may be contribute to regulation of glucokinase mRNA expression.

ISD 모델에 기초한 농업인 안전교육 프로그램 개발

김진모,이택선,이경숙,김경란,길대환,김효철,이찬 한국농업교육학회 2007 농업교육과 인적자원개발 Vol.39 No.3

이 연구의 목적은 ISD 모델에 기초하여 농업인 안전교육 프로그램을 개발하는데 있으며,이를 위해 분석-설계-개발-실행-평가의 다섯 단계로 구성된 개발모형을 설정하였다. 구체적인 프로그램 내용을 개발하기 위해 환경분석,학습자분석,과제분석을 실시하였고,그 결과를 반영하여 ‘안전의식 함양’,‘근골격계질환’,‘농약중독예방’,‘농기계 안전 사용’의 4개 모듈과 10개의 레슨을 설계하였다. 요구분석 및 설계 결과를 바탕으로 농업인 안전교육 프로그램의 교재/보조 자료/매체를 개발하였다. 교재의 경우,학습자용과 강사용을 구분하여 개발하였다. 개발된 프로그램 및 운영체제에 대한 검토 및 보완을 위한 전문가 협의회를 개최하여 프로그램 시연회를 실시한 후 교육프로그램을 수정,보완하였다. 그 후,농작업안전시범마을의 농업인을 대상으로 농업인 안전교육 프로그램을 시험 운영하였고,그 결과를 반영하여 농업인 안전교육 프로그램을 완성하였다. 또한 이상의 프로그램 개발 과정을 통해 나타난 논점을 토대로 향후 교육 프로그램 개발을 위한 시사점을 제공하였다. The purpose of this study was to develop a agricultural safety and health training program based on ISD Model. We set up 5step(Analysis, Design, Development, Implementation, Evaluation) model for developing the program. After analyzing environment, learner and task, four modules(① Safety and Health Mindset, ② Musculoskeletal System Disorders, ③ Agrichemical Poisoning Prevention, and ③ Using Farm Machines Safely) and 10 lessons were designed. We developed teaching materials and supplement materials for the Agricultural Safety and Health Training Program based on need analysis and program design. Teaching materials consisted of the learner's and instructor's. The developed program was examined by expert groups and the weak points were revised through professional workshop. Finally, we tested the Agricultural Safety and Health Training Program at agricultural safety and health model village and fixed the program by reflecting the test results.

Cyclosporin A 로 유도된 생쥐 림프절의 세포성 면역억제에 관한 면역조직화학적 연구 : T 림프구, IL-2 수용기 및 NK세포의 변화를 중심으로 Based on the change of T lymphocytes, Il-2 receptors, and NK cells

김진택,박인식,안상현,최난희,김동환 동국대학교 한의학연구소 1998 東國韓醫學硏究所論文集 Vol.6 No.2

본 실험은 cyclosporin A(CsA)에 의한 시간의 경과에 따른 림프절에서의 세포성 면역억제를 조사하기 위해서 시행된 것으로 BALB/C계 생쥐에 10일동안 CsA(45mg/kg/day) 투여 후 림프절에서의 T 림프구, IL-2 수용기 그리고 자연살해(NK)세포의 분포 변화를 관찰하였다. 대조군의 림프절에서는 L3T4(CD4)에 양성반응을 보이는 도움 T림프구, Ly2(CD8)에 양성반응을 보이는 세포독성 T 림프구 그리고 CD25R에 양성반응을 보이는 IL-2 수용기를 가진 세포는 곁피질(paracortex)과 수질동(medullary sinus)에서 분포하였다. CsA투여 후 처음 3일까지는 이들 양성반웅세포의 분포 변화는 없었으며 양성반응성의 변화도 없었다. 그러나 CsA 투여 7일부터 양성반응 세포수의 감소와 양성반응성의 약화가 관찰되기 시작하였으며 이러한 변화는 시간이 경과하여 14일에 이르렀을 때 가장 큰 감소추세로 나타났다. 한편 NKl.1(CD56)에 양성반응을 보이는 자연살해세포는 피질과 수질에 분포하였으며 CsA 투여 후 시간의 경과에 따라 양성반응 세포수가 감소하였으며, 이러한 감소는 14일에서 가장 큰 것으로 나타났다. 이상의 결과로 미루어보아 CsA 투여는 림프절에서의 IL-2 분비저해를 통해 T 림프구와 NK세포의 활성을 차단하여 선택적이면서 효과적인 세포성 면역억제작용을 하고 있는 것으로 사료된다. Cyclosporin A(CsA) is a selective immunosuppressive agent that has been credited with improved survival of solid organ allografts. Lymph node of BALB/C mouse administered CsA immunohistochemically observed to understand immunosuppressive effects of CsA on T lymphocytes, IL-2 receptors, and natural killer NK cells in lymph node. CsA orally administered daily for 10days at the dose 4smg/kg/day/. The lymph node were obtained at day 3, 7, and 14 after CsA administration and embeded with paraffin, and then stained by following ABC method that used monoclonal antibody including L3T4(CD4), Ly2(CD8), IL-2R(CD25), and NK-1.1(CD56). There were little changes of reactive degree and number of helper T lymphocytes, cytotoxic T lymphocytes, IL.-2 receptors, and NK cells at day 3 after CsA administration, but they began to decrease at day 7. These decrease were greatest at day 14. The helper T lymphocytes, cytotoxic T lymphocytes, IL-2 receptors, and NK cells distributed in paracortex and medullary sinus. These results indicated that the secretion of IL-2 began to decrease at day 7 after CsA administration and subsequently to suppress T lymphocytes and NK cell as components of cell-mediated immunity.

白朮散과 白扁豆가 白鼠 小腸과 大腸 杯狀細胞內 粘液質에 미치는 影響

金鎭澤,金兌勳,金東煥 동국대학교 한의학연구소 1993 東國韓醫學硏究所論文集 Vol.2 No.2

This study was performed to identify the effect of Backchulsan and Backpeundu, on the mucosubstances of the goblet cells in the small and large intestines of the mouse. After injected castor oil into oral cavity, the experimental animals were administrated with 5.952mg per 30g body weight of Backchulsan, with 0.519mg per 309 body weight of Bckpeundu. The specimens used in this study were obtained from the small and large intestines of the mouse. All specimens were stained with periodic acid-Schiff reaction, alcian blue pH 2.5 and alcian blue pH 1.0 procedures. The results were as follows 1. The most remarkable decrease of neutral and acid mucosubstance was observed in the control group 1 and 6 hours after castor oil administration. 2. Increase of neutral and mucosubstance was observed in Backchulsan administration group 3 hours in comparision with that of control group administrated of castor oil, especially of the upper and lower villi regions. 3. Increase of neutral and acid rnucosubstance was observed in Backpeundu administration group 6 hours in comparision with that of control group administration of castor oil. 4. The most remarkable increase of neutral and acid mucosubstance was observed in Backchulsan administration group after 3 hours in comparision with that of Backpeundu administration group.