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.

백서의 실험적 치아이동시 connexin 43의 발현에 관한 연구

임정현,강경화,이종진,김은철,김상철 대한치과교정학회 2001 대한치과교정학회지 Vol.31 No.5

치아이동시 골세포간 활성 조화에서 세포간 교통의 중요성이 차츰 인식되고 있으며 세포간 교통을 위한 교통반의 존재가 확인되고 있다. 동물실험 모델에서 실험적 치아이동을 통해 조성된 압박 및 견인측 치주인대에서 교통반 단백의 일종인 connexin 43의 발현을 관찰함으로써 인접 연조직 및 골 조직 내에서 세포 신호 전달 양상의 한 부분을 파악하고자 하였다. Sprague-Dawley계 백서 27마리를 대조군(3마리)과 실험군(24마리)으로 나누었으며, 실험군은 견인력(75g)을 가한 후 12시간, 1일, 4일, 7일, 14일, 28일이 경과한 후 각각 4마리씩 희생시켜, connexin 43의 발현을 면역조직화학적으로 관찰한 바 다음과 같은 결과를 얻었다. 1.대조군에서의 connexin 43 발현은 치은, 상아질, 치주인대, 치수, 골세포에서 경미하였다. 2.교정적 치아이동에 반응하는 치수, 치주인대, 골모세포, 파골세포, 골세포부위에서 connexin 43의 발현이 많았으나 치은, 상아질, 상아모세포에서의 발현은 견인력 적용기간에 관계없이 대조군과 큰 차이가 없었다. 3.치수에서 connexin 43의 발현은 교정력을 가한 4일에서부터 증가하여 7일째까지 크게 증가한 후 14일째부터 감소되었으며 28일째에는 대조군과 차이가 없었다. 4.치주인대에서 connexin 43의 발현은 주로 혈관을 중심으로 치조골 면을 따라 압박측에서 주로 나타났는데 교정력을 가한 12시간에서부터 증가하여 4일에서 가장 많은 발현을 보인 후 7일부터 감소하기 시작하였다. 5.골모세포 및 골세포에서 발현은 실험 1일째부터 출현해 4일째에 가장 많은 발현을 보였고 7일 이후에는 크게 감소하였으며 파골세포에서의 발현은 골모세포와 골세포보다 실험 4일째에 더 많이 관찰할 수 있었다. Bone remodeling in response to force requires coordinated actions of osteoblasts, osteoclasts, osteocytes, and periodontal ligament cells. Coordination among these cells may be mediated, in part, by cell-to-cell communication via gap junctions. This study was designed to evaluate the expression of gap junction, connection 43 in periodontal tissue during the experimental movement of rat's incisors, by LSAB(labelled streptavidine biotin) immunohistochemical staining fort connexin 43. Twenty seven Sprague-Dawley rats were divided into a control group(3 rats), and 6 experimental groups(24 rats) where 75g of force was applied from helical springs across the maxillary incisors. Rats of experimental groups were sacrificed at 12 hours, 1, 4, 7, 14 and 28 days after force application, respectively. And the tissues of a control group and experimental groups were studied immunohistochemically. The results were as follows : 1.In control group, the expression of connexin 43 was rare in gingiva, dentin, cementum, periodontal ligament and bone cells. 2.In experimental group, the expression of connexin 43 was increased in pulp, periodontal ligament, osteoblasts, and osteoclasts, comparing to that in control. And it was rare in gingiva, dentin, and odontoblasts regardless of the duration of force application, which was not different from that of control group. 3.The expression of connexin 43 in pulp of experimental group began to increase in 4-day after force application and got to the highest degree at 7-day. And it decreased after 14-day to be similar to that of control group at 28-day. 4.The expression of connexin 43 in periodontal ligament was noted in small capillaries adjacent to alveolar bone, showing higher intensity of immunolabelling after 4-day. And it was stronger in the pressure side than in tension side of periodontal ligament. After 7-day, decrease in connexin 43 expression was observed. 5.The expression of connexin 43 in alveolar bone began to increase 1-day, reached to the highest degree at 4-day, and decreased at 7-day. And the expression in osteoclasts was more than that in osteoblasts or osteocyte at 7-day.

위암의 진행에 따른 Fas관련 아포프토시스 유도의 상관성

임성철,황철기,오서진 조선대학교 부설 의학연구소 2000 The Medical Journal of Chosun University Vol.25 No.1

Background and Objectives : The purpose of this study is to determine whether human gastric adenocarcinomas express Fas-L, sFas-L or Fas , whether serum sFas-L leyel is changed in gastric cancer patients after gastrectomy, whether Fas-L expression is associated with increased apoptotic induction, especially, tumor-infiltrating Iymphocytes(TIL) and whether apoptotic induction is associated with the tumor stage and histologic type. Materials and Methods : The author analysed 38 cases of early gastric carcinoma(EGC) and 61 cases of advanced gastric carcinoma (AGC) which received gastric resection from 1997 to 1998. Of them, the number of diffuse type is 38 cases and the number of intestinal type is 61 cases. The author used immunohistochemical staining for Fas, Fas L and CD45, TACS^TM in situ apoptosis detection kit, and sFas ligand ELISA kit. Results : Fas-L was localized to neoplastic cells in 23 cases (23/38; 61%) of EGC group and 40cases (40/61; 66%) of AGC group. The extent of Fas-L expression was variable, with both Fas-L was localized to neoplastic cells in 23 cases (23/38; 61%) of EGC group and 40 cases(40/61; 66%) of group. The extent of Fas-L expression was variable, with both FasL-positive and negative neoplastic region cccuring within. TIL were detected by co-expression of CD4S and TACS on serial histologic sections. TIL adjacent to Fas-L erxressing tumor regions were decreased in number and TIL adjacent to FasL-negative tumor regions were increased in number; apoptotic induction of TIL showed the opposite pattern (p<05). Fas expression was found essentially homogeneously throughout the tumor mass independent of tumor stage. Fas expression showed 39 cases (39/61; 64%) of intestinal type and 26 cases (26/38; 68%) of diffuse type. Labeling indces for tumoral apoptosis in EGC and AGC were 6.72% and 7.13%, respectively and this difference was statistically insignificant. Co-expression of Fas-L and Fas, which occurred over large areas of the tumors, did not result in an enhanced rate of tumor cell apoptosis. The mean serum sFas-L level was significantly higher in patients before treatment compared with controls, whereas in post-gastrectomy patients, it was significantly lower, In addition, tumor stage and other prognostic factors were not associated with Fas and Fas-L expression, serum sFas-L level, number of TIL and apoptotic induction. Conclusion: 'The author demonstrates a statistically significant reduction of TIL concomitant with significantly increased TIL apoptosis adjacent to FasL-expressing regions of gastric adenocarcinomas. Also an elevated level of ·serum sFas-L in the gastric adenocarcinoma group was noted. These findings suggest Fas-mediated apoptotic depletion of TIL in response to Fas-L expression by stomach cancers, and provide the evidence to support the Fas counter-attack as a mechanism of immune escape in gastric cancer. And These findings indicate that the serum sFas-L level is a useful indicator in evaluating postoperative follow-up. In addition, gastric carcinoma cells of the intestinal and diffuse type did not differ in their expression of the Fas-apoptotic system.

혼수상태의 외상환자에서 초음파를 이용한 흉부 및 복부손상의 진단

임경수,이강현,이진웅,이부수,황성오,유수영,강성준 대한응급의학회 1995 대한응급의학회지 Vol.6 No.1

The need for rapid diagnosis and treatment of life-threatening thoracic and intra-abdominal injury result in controversy over the appropriate triage of unconscious blunt trauma patients. To aid in early decisions for these patients, a prospective analysis of 98 patients with glasgow coma scale(GCS) scores≤8 was undertaken. Although intra-thoracic and intra-abdominal injuries were frequently identified based on systolic blood ressure, the use of clinical signs alone resulted in more missed injuries than did using the emergency ultrasonography. In normotensive patients(n=34), intra-thoracic injuries was identified in one patient(2.9%), and intra-abdominal injuries were in 7 patients(20.6%). In shock(systolic blood pressure<90mmHg) Pa-tients(n=64), thoracic injuries and intra-abdominal injuries were diagnosed in 8(12.5%) and 27 patients(42.3%), thoracic and abdominal combined injuries were identified in 11 patients(17.2%). The sensitivity, specificity, and accuracy of emergency ultrasonography were 96.3%, 91.7%, 94.9%. This study suggests that all unconscious trauma patients undergo immediate emergency ultrasonography to prevent missing life-threatening injuries.

심정지환자에서 epinephrine투여시 혈중 catecholamine의 변화

임경수,황성오,이부수,이강현,이진웅,임종천,김영식,김선만,김 현 대한응급의학회 1995 대한응급의학회지 Vol.6 No.1

Background : Cardiac arrest is a potent stimulus for the release of endogenous catecholamine and high plasma catecholamine concentration has been reported during cardiopulmonary resuscitation(CPR). Few data were present about the clinical effect of high plasma catecholamine and endogenous catecholamine response to exogenous epinephrine administration during CPR. Purpose : This study was designed to evaluate endogenous catecholamine response to cardiac arrest and exogenously administered epinephrine during CPR. Method : In 21 cardiac arrest victims, blood sample for epinephrine and norepinephrine was drawn before and 1 minute after each epinephrine administration during CPR. 1mg of epinephrine was administered immediately after intravenous access and 3 minute interval during CPR. Plasma catecholamines were quantitated by high-performance liquid chromatography. Result : Baseline plasma epinephrine concentraqtion was higher in patients with cardiac arrest than normal controls (985±1627 vs 194±173 pg/ml, p<0.05). Baseline plasma norepinephrine concentration was also elevated in patients with cardiac arrest than normal controls, although statistically insignificnat(1965±4915 vs 360±250 pg/ml). Plasma epinephrine concentration in cardiac arrest patients was significantly elevated after exogenous administration of epinephrine, but its response was blunted after the third does of epinephrine. Plasma norepinephrine concentration in cardiac arrest patients was also elevated, but its response was observed only after the first epinephrine administration. Plasma epinephrine concentration was persistently higher in patients with return of spontaneous circulation (ROSC) than patients without ROSC. there was no significant difference of plasma norepinephrine concentration whether spontaneous circulation was restored or not. Endogenous epinephrine response to exogenous epinephrine was not observed in patients having prolonged arrest time over 20 minutes. Conclusion : Results of this study suggests that significant amount of catecholamine is released by exogenous epinephrine administration in spite of high plasma catecholamine concentration, and degree of endogenous catecholamine response influences resuscitation outcome.

인체 유방암종에서 Cyclic-Dependent Kinase Inhibitor, p27^Kip1 발현의 의의

송상용,김덕환,서연림,고영혜,김대식,남석진,양정현 대한병리학회 1998 Journal of Pathology and Translational Medicine Vol.32 No.12

농흉과 종격동염이 합병된 viridans streptococcus에 의한 인후농양 1예

임상현,유진홍,권순석,김연식,신완식,강문원,정명희 대한화학요법학회 1997 대한화학요법학회지 Vol.15 No.2

최근 저자들은 viridans streptococcus에 의한 인후농약으로 인하여 발생한 종격동염 및 농흉을 보인 47세 여자 환자를 적절한 배농과 항생제로 치료한 국내 최초의 1예를 치험하였기에 문헌 고찰과 함께 보고하는 바이다. Since the antibiotic era, retropharyngeal abscess became a rare disease entity nowadays. But it may often progress to lethal complications involving vital structure if diagnosis or treatment is delayed. We experienced a case of empyema and mediastinitis complicating retropharyngeal abscess caused by viridans streptococcus in a 47-year-old female patient with diabetes mellitus. She was successfully treated with appropriate antibiotic therapy combined with percutaneous abscess drainage. To our knowledge, such a complication of retropharyngeal abscess has not been reported in Korea yet and we present this unusual case herein followed by a review of the related literature.

치아우식증 수복용 북합레진의 개발과 치아 적합성에 관한 연구 제1편 : 치과용 복합레진의 개발과 기계적 성질의 평가에 관한 연구

임호남,박상진,김형찬 大韓齒科器材學會 1992 대한치과재료학회지 Vol.19 No.2

An esthetic and permanent restorative materials that could be placed directly into the cavities prepared in anterior teeth would be an important contribution to dental practice and public well-being. This studies were planned to produce some experimental composite resins with the evaluation of literatures related to manufacture processes of it. It was assumed that through this studies we wish to get some fundamental data using for the domestic production of commercial dental composite resins with the confirmation of manufacturing process repoted by forein researchs, and to evaluate the physical properties of it. Summarized manufacturing process used in this studies include the synthesis of BIS-GMA inlaboratory, the silanization of fillers, and the its incorporation into prepared BIS-GMA resin to make two paste system. As a first research project, its usefulness as a composite resin and characteristics under the some experimental conditions were evaluated by the measurements of hardening time, compressive strength, diametral tensile strength, and elastic modulus, and the observation of surface using SEM. Following conclusions were drawn: 1. Concentration control of initiator and accelerator from 2% to 4% resulted the working time ranged from 0.33 minutes to 1.00 minute and the setting time from 1.66 minute to 5 minutes. 2. Compressive strength of unfilled resins was highest on KM-33 specimens, diametral tensile strength were decreased by rise of accelerator contents, but it was increased by the rised content of initiator. 3. Higher storage temperature made all of experimental resins became excellent ones. 4. By the incorporation of filler, resins become harder and stronger with the higher values of measurement, but it was decreased by the long term immersion in water. 5. Both of filled and unfilled resins showed the higher dimetral tensile strength over each allowable values of American Dental Specification.

염농도에 따른 멸치젖의 품질 변화

임상빈,오창경,송대진 濟州大學校 産業技術硏究所 2000 산업기술연구소논문집 Vol.11 No.1

General composition, pH. salinity. peroxide value. TBA value. volatile basic nitrogen(VBN) and amino nitrogen were measured for fermented anchovy with different salt content. Moisture. crude protein, crude lipid and ash of raw anchovy were 75.9, 18.2, 2.36 and 3.9%. respectively. Moisture of fermented anchovy decreased as the increase of salt added. Crude protein and crude lipid were the highest in 12% salt fermented anchovy. Ash of fermented anchovy increased as the increase of salt content. pH and salinity of raw anchovy were 7.0 and 0.61%. respectively. pH of fermented anchovy were 6.0 regardless of salt concentration. Peroxide values increased drastically with the increase of salt added. TBA was the lowest and amino nitrogen was the highest in 8% salt fermented anchovy.

인체 체액내 황색포도상구균의 증식과 철 획득 기전

신성희,이강길,이명희,양남웅,임용,김진호 대한감염학회 2000 감염 Vol.32 No.5