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Transfer of Isolated Mitochondria to Bovine Oocytes by Microinjection (미세주입을 이용한 난자로의 분리된 미토콘드리아 전달)

  • Baek, Sang-Ki;Byun, June-Ho;Kim, Bo Gyu;Lee, A ram;Cho, Young-Soo;Kim, Ik-Sung;Seo, Gang-Mi;Chung, Se-Kyo;Lee, Joon-Hee;Woo, Dong Kyun
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1445-1451
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    • 2017
  • Mitochondria play a central role in energy generation by using electron transport coupled with oxidative phosphorylation. They also participate in other important cellular functions including metabolism, apoptosis, signaling, and reactive oxygen species production. Therefore, mitochondrial dysfunction is known to contribute to a variety of human diseases. Furthermore, there are various inherited diseases of energy metabolism due to mitochondrial DNA (mtDNA) mutations. Unfortunately, therapeutic options for these inherited mtDNA diseases are extremely limited. In this regard, mitochondrial replacement techniques are taking on increased importance in developing a clinical approach to inherited mtDNA diseases. In this study, green fluorescence protein (GFP)-tagged mitochondria were isolated by differential centrifugation from a mammalian cell line. Using microinjection technique, the isolated GFP-tagged mitochondria were then transferred to bovine oocytes that were triggered for early development. During the early developmental period from bovine oocytes to blastocysts, the transferred mitochondria were observed using fluorescent microscopy. The microinjected mitochondria were dispersed rapidly into the cytoplasm of oocytes and were passed down to subsequent cells of 2-cell, 4-cell, 8-cell, morula, and blastocyst stages. Together, these results demonstrate a successful in vitro transfer of isolated mitochondria to oocytes and provide a model for mitochondrial replacement implicated in inherited mtDNA diseases and animal cloning.

EVALUATION ON THE ABRASION RESISTANCE OF A SURFACE SEALANT (레진전색제의 마모저항성에 대한 평가)

  • Kim, Soo-Mee;Han, Sae-Hee;Cho, Young-Gon
    • Restorative Dentistry and Endodontics
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    • v.32 no.3
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    • pp.180-190
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    • 2007
  • The purpose of this study was to evaluate the abrasion resistance of surface penetrating sealant which was applied on a composite resin restoration and to provide proper time to reapply sealant on composite resin surface. Two hundred rectangular specimens, sized $8\times3\times2mm$, were made of Micronew (Bisco, Inc., Schaumburg, IL, U.S.A) and divided into two groups; F group (n = 10) was finished with coarse and medium grit of Sof-Lex discs and BisCoverwas applied B group (n = 190) after finishing with discs. B group was again subdivided into nineteen subgroups From B-1 group to B-18 group were subjected to toothbrush abrasion test using a distilled water-dentifrice slurry and toothbrush heads B-IM group was not subjected to toothbrush abrasion test. Average surface roughness (Ra) of each group was calculated using a surface roughness tester (Surfcorder MSE-1700: Kosaka Laboratory Ltd., Tokyo, Japan) . A representative specimen of each group was examined by FE-SEM (S-4700: Hitachi High Technologies Co., Tokyo, Japan). The data were analysed using cluster analysis, paired t-test, and repeated measure ANOVA. The results of this study were as follows; 1. Ra off group was $0.898{\pm}0.145{\mu}m$ and B-IM group was $0.289{\pm}0.142{\mu}m$. Ra became higher from B-1 group $(0.299{\pm}0.48{\mu}m$ to B-18 group $(0.642{\pm}0.313{\mu}m$. 2. Final cluster center of Ra was $0.361{\mu}m$ in cluster 1 $(B-IM\simB-7)$, $0.511{\mu}m$ in cluster 2 $(B-8\simB-14)$ and $0.624{\mu}m$ in cluster 3 ($(B-15\simB-18)$. There were significant difference among Ra of three clusters. 3 Ra of B-IM group was decreased 210.72% than Ra of F group. Ra of B-8 group and B-15 group was increased 35.49% and 51.35% respectively than Ra of B-IM group. 4. On FE-SEM, B-IM group showed the smoothest resin surface. B-8 group and B-15 group showed vertically shallow scratches , and wide and irregular vertical scratches on composite resin surface respectively. Within a limitation of this study, finished resin surface will be again smooth and glazy if BisCover would be reapplied within 8 to 14 months after applying to resin surface.

Radiation Therapy for Pituitary Adenoma -Changes in Endocrine Function after Treatment- (뇌하수체선종의 방사선치료후 혈중 호르몬치의 변화)

  • Yoon Sei Chul;Jang Hong Suck;Kim Song Hwan;Shinn Kyung Sub;Bahk Yong Whee;Son Ho Young;Kang Joon Ki
    • Radiation Oncology Journal
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    • v.9 no.2
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    • pp.185-195
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    • 1991
  • Seventy four patients with pituitary adenoma received radiation therapy (RT) on the pituitary area using 6 MV linear accelerator during the past 7 years at the Division of Radiation Therapy, Kangnam St. Mary's Hospital, Catholic University Medical College. Thirty nine were men and 35 were women. The age ranged from 7 to 65 years with the mean being 37 years. Sixty five ($88\%$) patients were treated postoperatively and 9 ($12\%$) primary RT, To evaluate the effects of RT, we analyzed the series of endocrinologic studies with prolactin (PRL), growth hormone (GH), adrenocorticotrophic hormone (ACTH), leuteinizing hormone (LH), follicular stimulating hormone (FSH) and thyroid stimulating hormone (TSH) etc after RT. All but one with Nelson's syndrome showed abnormal neuroradiologic changes in the sella turcica with invasive tumor mass around supra- and/or parasella area. The patients were classified as 23 ($29\%$) prolactinomas and 20 ($26\%$) growth hormone (GH) secreting tumors, and 6 ($8\%$ ACTH secreting ones consisting of 4 Cushing's disease and 2 Nelson's syndrome. Twentynine ($37\%$) had nonfunctioning tumor and four ($5\%$) of those secreting pituitary tumors were mixed PRL-GH secreting tumors. The hormonal level in 15 ($65\%$) of 23 PRL and 3 ($15\%$) of 20 GH secreting tumors returned to normal by 2 to 3 years after RT, but five PRL and five GH secreting tumors showed high hormonal level requiring bromocriptine medication. Endocrinologic insufficiency developed by 3 years after RT in 5 of 7 panhypopituitarisms, 4 of seven hypothyroidisms and one of two hypogonadisms, respectively. Fifteen ($20\%$) patients were lost to follow up after RT.

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Analysis of Treatment Failure after Curative Radiotherapy in Uterine Cervical Carcinoma (자궁경부암에 있어서 방사선치료 후의 치료실패 분석)

  • Chai, Gyu-Young;Kang, Ki-Mun;Lee, Jong-Hak
    • Radiation Oncology Journal
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    • v.19 no.3
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    • pp.224-229
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    • 2001
  • Purpose : The aim of this study is to analyze the treatment failure patterns and the risk factors for locoregional or distant failure of uterine cervical carcinoma treated with radiation therapy. Materials and methods . A retrospective analysis was undertaken of 154 patients treated with curative radiation therapy in Gyeongsang National University Hospital from April 1989 through December 1997. According to FIGO classification, 12 patients were stage IB, 24 were IIA, 98 were IIB, 1 were IIIA, 17 were IIIB, 2 were IVA. Results : Overall treatment failure rate was $42.1\%$ (65/154), and that of complete responder was $31.5\%$ (41/130). Among 65 failures, 25 failed locoregionally, another 25 failed distantly, and 15 failed locoregionally and distantly. Multivariate analysis confirmed tumor size (>4 cm) as risk factor for locoregional failure, and tumor size (>4 cm), pelvic lymph node involvement as risk factors for distant failure. Conclusion : On the basis of results of our study and recent published data of prospective randomized study for locally advanced uterine cervical carcinoma, we concluded that uterine cervical carcinoma with size more than 4 cm or pelvic lymph node involvement should be treated with concurrent chemoradiation.

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Diagnostic Methods Used in the Bone Infections in Children (소아기 골 감염의 진단방법에 관한 조사)

  • Lee, Eun Sil;Choi, Kwang Hae
    • Pediatric Infection and Vaccine
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    • v.4 no.2
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    • pp.210-217
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    • 1997
  • Purpose: To prevent residual physical disability and chronic infection, prompt diagnosis and adequate treatment are important in the skeletal infections in children. Although radioisotope scanning is knwon as the method of choice for early diagnosis of bone infection, we conducted a study on twenty nine children who had skeletal infections to reevaluate the most appropriate way in diagnosis and management. Methods: A retrospective study was conducted on twenty nine children, who were admitted to the departments of Pediatrics and Orthopedic Surgery and who had acute osteomyelitis or septic arthritis, through review of medical records, radiologic & radioisotope study results. Their diagnoses were confirmed by bacteriologic cultures on the aspirated specimens from suspected bony lesions. Results: 1) Among twenty nine patients, there were 6 infants including 5 newborn infants, and 23 children were aged between 1 and 15 years. Male to female ratio was 1.4 to 1. 2) Point tenderness was noted in all cases, and the common physical signs were swelling, limitation of motion, fever and local heat in the order of frequency. 3) Fifty two percents of the patients were diagnosed within a week after onset of symptoms and all cases were within 15 days. 4) Leukocytosis was noted in only 58.6% of cases but erythrocyte sedimentation rate was increased in all cases except only one case. Staphylococcus aureus was revealed as the most common etiologic agent. 5) Radioisotope scans showed hot uptake in five of six cases(83.3%) who had no abnormal finding on plain skeletal radiolograms. Conclusions: Although radioisotope scan and MRI are helpful in early diagnosis before radiologic finding was detected on plain X-ray film, the antimicrobial therapy can be started after bacteriologic study of the aspirated specimens from the suspected skeletal lesions if skeletal infection is highly suspected clinically.

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The effect of tumor necrosis factor (TNF)-α to induce matrix metalloproteinase (MMPs) from the human dental pulp, gingival, and periodontal ligament cells (사람의 치수, 치은, 치주인대 세포에 tumor necrosis factor (TNF)-α로 자극 시 matrix metalloproteinase (MMPs)의 분비에 관한 연구)

  • Rhim, Eun-Mi;Park, Sang-Hyuk;Kim, Duck-Su;Kim, Sun-Young;Choi, Kyoung-Kyu;Choi, Gi-Woon
    • Restorative Dentistry and Endodontics
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    • v.36 no.1
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    • pp.26-36
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    • 2011
  • Objectives: In the present study, three kinds of tissues cells (pulp, gingiva, and periodontal ligament) were investigated if those cells express MMP and TIMP when they were stimulated with neuropeptides (substance P, CGRP) or proinflammatory cytokine, TNF-$\alpha$. Materials and Methods: The cells cultured from human dental pulp (PF), gingiva (GF) and periodontal ligament were (PDLF) stimulated with Mock, SP, TNF-$\alpha$, and CGRP for 24 hrs and 48 hrs. for an RNase protection assay and Enzyme Linked Immunosorbent Assay. Cells (PF, GF and PDLF) seeded in 100 mm culture dish were stimulated with SP ($10^{-5}$, $10^{-8}\;M$) or only with medium (Mock stimulation) for 4hrs and for 24 hrs for RNase Protection Assay, and they were stimulated with CGRP ($10^{-5}\;M$) and TNF-$\alpha$(2 ng/mL) for 24 hrs and with various concentraion of TNF-$\alpha$(2, 10, and 100 ng/mL) for Rnase Protection Assay with a human MMP-1 probe set including MMP 1, 2, 8, 7, 8, 9, 12, and TIMP 2, 3. In addition, cells (PF, GF and PDLF) were stimulated with Mock and various concentraion of TNF-$\alpha$(2, 10, and 100 ng/mL) for 24 hrs and with TNF-$\alpha$(10 ng/mL) for 48 hrs, and the supernatents from the cells were collected for Enzyme Linked Immunosorbent Assay (ELISA) for MMP-1 and MMP-13. Results: The expression of MMPs in PF, GF, PDLF after stimulation with SP and CGRP were not changed compared with Mock stimulation for 4 hrs and 24 hrs. The expression of MMP-1, -12, -13 24 hrs after stimulation with TNF-$\alpha$ were upregulated, however the expression of TIMP-3 in PF, GF, PDLF after stimulation with TNF-$\alpha$ were downregulated. TNF-$\alpha$(2 ng/mL, 10 ng/mL, 100 ng/mL) increased MMP-1 and MMP-12 expression in PF dose dependently for 24 hrs. Conclusions: TNF-$\alpha$ in the area of inflammation may play an important role in regulating the remodeling of dentin, cementum, and alveolar bone.

The evaluation of color and color difference according to the layering placement of Incisal shade composites on the body composites of the indirect resin restoration (간접 수복용 복합레진의 Incisal 색상 적층 두께에 따른 표면 색상 및 색차의 평가)

  • Park, Su-Jung;Lee, Han-Young;Nah, Myong-Yun;Chang, Hoon-Sang;Hwang, Yun-Chan;Oh, Won-Mann;Hwang, In-Nam
    • Restorative Dentistry and Endodontics
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    • v.36 no.1
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    • pp.37-49
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    • 2011
  • Objectives: The aim of this study was to evaluate the surface color of indirect resin restoration according to the layering placement of different shade of incisal composite. Materials and Methods: In this study, CIE $L^*a^*b^*$ value of 16 Body composite of Tescera ATL (Bisco, Schaumburg IL,USA) was measured by spectrophotometer (NF999, Nippon Denshuku, Japan), and compared to CIE $L^*a^*b^*$ value of Vitapan shade guide. Nine shade Incisal composite of Tescera ATL were buildup to 1 mm thickness on Body composites inlay block, and CIE $L^*a^*b^*$ value was measured. Incisal composite was ground to 0.5 mm thickness and CIE $L^*a^*b^*$ value was re-measured. Color difference between Body composite and Incisal composites layered on Body composite was calculated as a function of thickness. Results: Color difference between corresponding shade of Tescera Body composite and Vitapan shade guide was from 6.88 to 12.80. $L^*$ and $b^*$ value was decreased as layering thickness of Incisal composite on Body composite was increased. But, $a^*$ value did not show specific change tendency. Conclusions: Surface color difference between Body composites and Incisal composites layered on Body composite was increased as the layering thickness of Incisal composite increased (p < 0.05).

The Association between Bone Mineral Density, Bone Turnover Markers, and Nutrient Intake in Pre- and Postmenopausal Women (폐경 전.후 여성의 골밀도 및 골대사 지표에 영향을 미치는 요인)

  • Park, Ji-Youn;Choi, Mi-Youn;Lee, Seon-Heui;Choi, Yoon-Ho;Park, Yoo-Kyoung
    • Journal of Nutrition and Health
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    • v.44 no.1
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    • pp.29-40
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    • 2011
  • The purpose of this study was to examine the association among bone mineral density (BMD), biochemical bone markers, nutrients, and salt intake in premenopausal and postmenopausal women. We evaluated 431 subjects who visited a health promotion center of a university hospital between January 2008 and July 2009. We excluded those who were taking medications or who had an endocrine disorder affecting osteoporosis. The subjects were divided into premenopausal (n = 283) and postmenopausal (n = 143) women. We evaluated the correlation among BMD of the lumbar spine, femoral neck, and total femoral, as well as biochemical bone markers, hormone, serum profiles, general characteristics, nutrient intakes, and food intake frequencies. From a stepwise multiple regression analysis, lumbar spine BMD was positively correlated with weight (p < 0.001) and negatively correlated with osteocalcin (OC)(p < 0.001), Femoral neck BMD was positively correlated with weight (p < 0.001) and negatively correlated with C-telopeptide (CTx) and alkaline phosphatase (ALP)(p < 0.001, p < 0.05). In premenopausal women, femoral total BMD was positively correlated with BMI (p < 0.001) and negatively correlated with CTx (p < 0.001). In postmenopausal women, lumbar spine BMD was positively correlated with calcium intake (p < 0.01) and negatively correlated with sodium intake (p < 0.01). Femoral neck and femoral total BMD were both positively correlated with weight (p < 0.001), and femoral neck BMD was negatively correlated with age and ALP (p < 0.001, p < 0.05). Femoral total BMD was negatively correlated with age and OC (p < 0.001, p < 0.01). These results suggest that reducing sodium intake may play an important role delaying bone resorption and preventing a decrease in BMD.

Development of an accreditation system for dietary and nutrition related education resources (영양.식생활 교육자료의 인증 시스템 개발 연구)

  • Kim, Ji-Myung;Lee, Kyoung Ae;Park, Yoo Kyoung;Lee, Kyung-Hea;Oh, Sang Woo;Lee, Hee Seung
    • Journal of Nutrition and Health
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    • v.47 no.2
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    • pp.145-156
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    • 2014
  • Purpose: The purpose of this study was to establish accreditation systems of reliable educational materials for nutrition and dietary life which could be used in schools, workplace, and health promotion. Methods: The study was conducted from April 2011 to October 2011. Literature reviews, institutional visits, and telephone interviews were conducted. Expert meetings and advisory councils were held in order to receive feedback on development of the accreditation systems. A survey was conducted for the accreditation procedures on 143 professionals, including professors, researchers, health and medical experts, teachers, nutrition teachers, dietitians, and clinical nutritionists. Results: The final procedure of the developed accreditation system was finalized as follows: 1) receiving application twice per year 2) complete desk review (written evaluation) by three reviewers within two months, 3) board review (all board members) and decision, and 4) notification of results. The accreditation system is set for printed materials, web-site, and materials for activities. The certificate and accreditation mark is issued to the final certified educational materials. Expiration date is established only for the web-site form. The accreditation length lasts for two years, and can be extended by renewal application. Conclusion: The dietary and nutrition related materials, which are certificated by this accreditation system, could impart reliable information and knowledge to both learners and educators, and help them in effective selection of educational materials. Therefore, this accreditation system might be expected to increase satisfaction for teaching and learning about nutrition and healthy dietary life.

A Case of Late-onset Episodic Myopathic Form with Intermittent Rhabdomyolysis of Very-long-chain acyl-coenzyme A Dehydrogenase (VLCAD) Deficiency Diagnosed by Multigene Panel Sequencing (유전자패널 시퀀싱으로 진단된 성인형 very-long-chain acyl-coenzyme A dehydrogenase (VLCAD) 결핍증 증례)

  • Sohn, Young Bae;Ahn, Sunhyun;Jang, Ja-Hyun;Lee, Sae-Mi
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.19 no.1
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    • pp.20-25
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    • 2019
  • Very-long-chain acyl-CoA dehydrogenase (VLCAD) deficiency (OMIM#201475) is an autosomal recessively inherited metabolic disorder of mitochondrial long-chain fatty acid oxidation. The clinical features of VLCAD deficiency is classified by three clinical forms according to the severity. Here, we report a case of later-onset episodic myopathic form of VLCAD deficiency whose diagnosis was confirmed by plasma acylcarnitine analysis and" multigene panel multigene panel sequencing. A 34-year old female patient visited genetics clinic for genetic evaluation for history of recurrent myopathy with intermittent rhabdomyolysis. She suffered first episode of rhabdomyolysis with acute renal failure requiring hemodialysis at twelve years old. After then, she suffered several times of recurrent rhabdomyolysis provoked by prolonged exercise or fasting. Physical and neurologic exam was normal. Serum AST/ALT and creatinine kinase (CK) levels were mildly elevated. However, according to her previous medical records, her AST/ALT, CK were highly elevated when she had rhabdomyolysis. In suspicion of fatty acid oxidation disorder, multigene panel sequencing and plasma acylcarnitine analysis were performed in non-fasting, asymptomatic condition for the differential diagnosis. Plasma acylcarnitine analysis revealed elevated levels of C14:1 ($1.453{\mu}mol/L$; reference, 0.044-0.285), and C14:2 ($0.323{\mu}mol/L$; 0.032-0.301) and upper normal level of C14 ($0.841{\mu}mol/L$; 0.065 -0.920). Two heterozygous mutation in ACADVL were detected by multigene panel sequencing and confirmed by Sanger sequencing: c.[1202G>A(;) 1349G>A] (p.[(Ser 401Asn)(;)(Arg450His)]). Diagnosis of VLCAD deficiency was confirmed and frequent meal with low-fat diet was educated for preventing acute metabolic derangement. Fatty acid oxidation disorders have diagnostic challenges due to their intermittent clinical and laboratorial presentations, especially in milder late-onset forms. We suggest that multigene panel sequencing could be a useful diagnostic tool for the genetically and clinically heterogeneous fatty acid oxidation disorders.

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