• Title/Summary/Keyword: One stage fusion

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Western blot diagnosis of vivax malaria with multiple stage-specific antigens of the parasite

  • Son, Eui-Sun;Kim, Tong-Soo;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.39 no.2
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    • pp.171-176
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    • 2001
  • Western blot analysis was performed to diagnose vivax malaria using stage-specific recombinant antigens. Genomic DNA from the whole blood of a malaria patient was used as templates to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), meroxoite surface protein (MSP-1), apical merozoite antigen (AMA- 1), serine repeat antigen (SERA), and exported antigen (EXP- 1) of Plasmodium vivax. Each amplified DNA fragment was inserted into a pGEX-4T plasmid to induce the expression of GST fusion protein in Escherichia coli by IPTG. The bacterial cell extracts were separated on 10% SDS-PAGE followed by western blot analysis with patient sera which was confirmed by blood smear examination. When applied with patient sera, 147 (91.9%) out of 160 vivax malaria, 12 (92.3%) out of 13 falciparum malaria, and all 9 vivax/falciparum mixed malaria reacted with at least one antigen, while no reactions occurred with 20 normal uninfected sera. In the case of vivax malaria, CSP-1 reacted with 128 (80.0%) sera, MSP-1 with 102 (63.8%), AMA-1 with 128 (80.0%), SERA with 115 (71.9%), and EXP-1 with 89 (55.6%), respectively. We obtained higher detection rates when using S antigens (91.9%) rather than using each antigen solely (55.6 - 80%), a combination of 2 (76.3 - 87.5%), 3 (85.6 - 90.6%), or 4 antigens (89.4 - 91.3%). This method can be applied to serological diagnosis, mass screening in endemic regions, or safety test in transfusion of prevalent vivax malaria.

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Generation of Embryonic Stem Cell-derived Transgenic Mice by Using Tetraploid Complementation

  • Park, S.M.;Song, S.J.;Uhm, S.J.;Cho, S.G.;Park, S.P.;Lim, J.H.;Lee, H.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.12
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    • pp.1641-1646
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    • 2004
  • The objective of this study was to generate transgenic mice expressing human resistin gene by using the tetraploidembryonic stem (ES) cell complementation method. Human resistin gene was amplified from human fetal liver cDNA library by PCR, cloned into $pCR^{(R)}$ 2.1 $TOPO^{(R)}$ vector and constructed in pCMV-Tag4C vector. Mammalian expression plasmid containing human resistin was transfected into D3-GL ES cells by Lipofectamine 2,000, and then after 10-12 days of transfection, the human resistin-expressing cells were selected with G418. In order to produce tetraploid embryos, blastomeres of diploid embryos at the two-cell stage were fused with two times of electric pulse using 60 V 30 $\mu$sec (fusion rate: 2,114/2,256, 93.5%) and cultured up to the blastocyst stage (development rate: 1,862/2,114, 94.6%). The selected 15-20 ES cells were injected into tetraploid blastocysts, and then transferred into the uteri of E 2.5 d pseudopregnant recipient mice. To investigate the gestation progress, two E 19.5 mused fetuses were recovered by Cesarean section of which one fetus was confirmed to contain human resistin gene by genomic DNA-PCR. Therefore, our findings demonstrate that tetraploid-ES mouse technology can be considered as a useful tool to produce transgenic mice for the rapid analysis of gene function in vivo.

Studies on the Cleavage Inhibidng Activity in the Cytoplasm of Growing Follicular Oocytes in Mammals (성장중인 포유동물 여포난자 세포질의 난할억제효과에 관하여)

  • 이원교;권혁방
    • The Korean Journal of Zoology
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    • v.33 no.1
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    • pp.45-52
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    • 1990
  • In order to determine whether maturation inhibiting activity(MIA) in the cytoplasm of growing follicular oocytes would suppress the cleavage of the embryonal cells, the growing oocytes were fused with the 2 or 4 cell blastomeres and then examined for the nuclear phase of the fused giant cells 24 hr after culture. A significant number of the giant cells(60%) composed of growing mouse oocyte and 2 cell mouse blastomere(1/2) in interphase has contained 2 nuclei 24 hr after culture and most of the giant cells (90%) composed of the growing oocyte and 4 cell blastomere(1/4) also contained 2 nuclei after culture. The unfused blastomeres or the isolated blastomeres cultured without fusion treatment cleaved one cell cycle under the same culture condition. In contrast, the nucleus of the growing oocytes was disintegrated and the chromosome condensed when fused with 2 cell blastomere in mitosis. The growing rat oocytes also suppressed the nuclear disintegration of the mouse embryonal cells during culture. The data presented here showed that MIA in the growing mammalian oocyte inhibited the cleavage of the embryonal cells in interphase stage, but not in milosis stage.

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Strain Improvement of the Genus Pleurotus by Protoplast Fusion (원형질체(原形質體) 융합(融合)에 의한 느타리버섯속(屬)의 품종개발(品種開發))

  • Yoo, Young-Bok;You, Chang-Hyun;Cha, Dong-Yeul
    • The Korean Journal of Mycology
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    • v.21 no.3
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    • pp.200-211
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    • 1993
  • Somatic hybrids of Pleurotus florida ASI 2016 and Pleurotus ostreatus ASI 2018 were obtained by protoplast fusion. The 40 fusants($P1{\sim}P40$) was examined for the yield on fermented and pasteurized rice straw in a tray. The carpophore yield of them were showed as the range of $27.0{\sim}155.2$, based on parental values of 100(ASI 2018), The pilei of fusants between orange white colored P. florida and dark grey colored P. ostreatus had mixed colors in the young stage. Other breeding programmes were performed to improve new varieties with high yield and good quality. A new oyster mushroom variety, Wonhyeongneutaribeosus(P72), was developed at the Agricultural Sciences Institute, Rural Development Administration in 1990. This P.florida-ostreatus-ostreatus hybrid P72 was selected from 38 protoplast fusion products($P41{\sim}P78$) between P.florida-ostreatus recombinant P5-M 43-arg rib and P. ostreatus ASI 2-13-0 2001-19-pro orn. The yield indexes of 38 hybrids ranged $40.5{\sim}152.7$ compared with the parental values of 100(ASI 2001). Hybrid P72 was characterized by the large fruiting bundle of semispherical shape with long stipe and by the small and circular pileus, resulting in lower harvesting cost. A significant increase in carpophore production was observed in somatic hybrids of protoplasts due to heterosis. A comparision of hybrid with parents P72 was made using isozyme analysis. The esterase banding patterns could be characterized by new bands in the hybrids. Seven fusion products of four crosses between P.florida ASI 2016 and P. ostreatus ASI 2018 were analysed with respect to the distribution of progenies and segregation of gene markers by random basidiospore analysis. Segregation of alleles should yield progeny of four genotypes in a Mendelian ratio of 1 : 1 : 1 : 1 for prototrophs, auxotrophs of one parental type, auxotrophs of the other parental type, and auxotrophic recombinants, respectively. However, five fusants of them did not detect one parental, P.ostreatus, type. Basidiospores could yield progeny of 16 genotypes in the cross of one of the recombinant P5-M43-arg $rib{\times}P. ostreatus$ ASI 2-13-pro orn but the segregants of three fusants were not detected clearly. The allele ratio of loci could be expected 1 : 1 : 1 : 1 for arg, rib, pro and orn. The ratio, however, would be changed to 4 : 1 : 1 : 1 with increasing proportion of argo In almost all the fusants, prototrophic recombinants were recovered in large numbers against auxotrophic markers. Parental genotypes were recovered with the recombinant progeny amounting to $38.68{\sim}99.56%$. The analysis provides proof of heterokaryosis and strong evidence for haploidy of vegetative nuclei, a sexual cycle consisting of nuclear fusion and meiosis.

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The Outcome of the Treatment of Lesser toe Brachymetatarsia - focus on complications - (소 족지 단중족증의 치료결과 - 합병증을 중심으로 -)

  • Lee, Ho-Seung;Yoon, Joon-O;Park, Soo-Sung;Kim, Eu-Gene
    • Journal of Korean Foot and Ankle Society
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    • v.7 no.1
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    • pp.13-20
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    • 2003
  • Purpose: We analysis the outcome and complications of treatment of lesser toe brachymetatarsia. Materials and Methods: We analysed 28 patients 35 cases of lesser toe brachymetatarsia. Mean post operative follow up period was 2 years 8 months. All of the patients were female and mean age at operation was 21 years old. 2 cases of third metatasal bone and 33 cases of fourth metatarsal bone were operated. 8 metatarsal bones were treated using one staged lengthening with tricortical bone graft and 27 metatarsal bones were treated using callotasis with monofixator. Results: The average amount of lengthening was 13.3mm(12mm-15mm) in one staged lengthening, while 14.4mm(4mm-23mm) in callotasis. Average percentile increase was 28.9%(26%-34%) in one staged lengthening and 32%(18%-46%) in callotasis. The average healing index of callotasis was 76 days/cm (41 days/cm-166 days/cm). Satisfied outcomes in 4 cases of 8 cases (50%) after one staged lengthening and 17 cases of 27 cases (63%) after callotasis. 6 complications in 4 cases were occurred after one staged lengthening; insufficient length gain in 3 cases, fracture on the junction of graft bone and metatarsal bone in 1 case, plantar bowing deformity in 1 case and bony fusion of metatarsophalangeal joint in 1 case. 17 complications in 10 patients were occurred after callotasis ; metatarsophalangeal joint stiffness in 8 cases, metatarsophalangeal joint subluxation in 2 cases, overlengthened metatarsal bone in 2 cases, tapering of callus in 1 case, fracture of callus in 1 case, premature consolidation of callus in 1 case, osteomyelitis of metataral head in 1 case and plantar bowing deformity in 1 case. Conclusion: Although one staged lengthening and gradual lengthening using callotasis are effective treatment for lesser toe brachymetatarsia, complications not rarely occured after lengthening. Insufficient lengthening are most common complication after one staged lengthening while metatarsophalangeal joint stiffness lire most common complication after callotasis.

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A Study on the Strategy for Enhancing the Service Export linked with Manufacturing Sector : focused on Stage System and Special Lighting Service (제조-서비스 연계형 수출상품화 모델 개발전략 - 무대장치 및 특수조명서비스 수출산업을 중심으로 -)

  • Park, Moon-Suh
    • International Commerce and Information Review
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    • v.10 no.4
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    • pp.457-491
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    • 2008
  • As stage equipment export markets along with special lighting service lack the attraction for already globally established businesses, such markets can be viewed as an advantageous opportunity for SMEs as in general. In reality, global businesses tend to focus on large construction projects and this indicates relatively less substantial markets such as stage equipment and special lighting service export are more suitable for SME businesses. However, possible problems may be recognized as following; doubtful capabilities by such businesses to join in the vast and competitive global market and pursue manufacturing and service based export. This point is also supported by the fact that such in general SME businesses have substantially less experience in exporting products and services abroad. Realizing the distinctive features of the Korean economy, it is unarguable that every sector and area of global market must be regarded and monitored closely. Hence, it can be argued that there is an imminent need for establishment of supportive institution to assist export process of combination of stage equipments and special lighting service. This study emphasizes the need to improve export process of stage equipments, special lighting services as well as other related products and services which have been focused in domestic market only until now. Further, it also analyzed the potential prospect of such direction reconciling current crisis our manufacturing industry is facing. Even though it maybe regarded as one of the niche market for export of Korea in the short term view, stage equipment and special lighting service industry may rapidly grow as the global cultural industries have grown along with the increase of national income earnings overall. Due to such advantageous features, it can be expected that such industries will show strong growth in the near future. After analyzing the fact that Korea's plants (eg. powerplants) export sector is at its boom, there is a need to transform stage equipment and special lighting service export market into a primary market from a secondary(niche) market for SMEs. This study is viewed from the Korean economic and export sector aspect in the aim of seeking a solution to conquest our realistic limit in our export sector by developing a suitable export model. There have been cases of very few attempts to expand abroad by SMEs who have failed miserably due to their failure to adapt to foreign culture, practice and languages as well as substantial lack in experience in export marketing. Despite this, neglecting our manufacturing industry as it is which is showing its limit and problems is out of option therefore, it is imminent that we come up with an effective measure to address this problem and service export can be suggested as one of them. This study reveals manufacturing-service export model of stage equipment and special lighting service and its related areas is recognized as a field with a very strong future and furthermore, it is expected to bring synergy effects in manufacturing and services sector as well. Further, the operation strategy contains combination, composition and fusion(convergence) of manufacturing and service sectors which could derive various of export products which displays greater success probability or this export model. The outcome of this research is expected to become a useful source for enterprises related to such industry which are seeking a possible global expansion. Furthermore, it is also expected to become a catalyst which fastens the process of global expansion and not only that, we are firmly assured that this study will become an opportunity to improve our current policies and institutions related to this area's export market.

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Expression Analysis of the Mx Gene and Its Genome Structure in Chickens

  • Yin, C.G.;Du, L.X.;Li, S.G.;Zhao, G.P.;Zhang, J.;Wei, C.H.;Xu, L.Y.;Liu, T.;Li, H.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.7
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    • pp.855-862
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    • 2010
  • Among the known interferon-induced antiviral mechanisms, the Mx pathway is one of the most powerful pathways. The Mx protein has direct antiviral activity and inhibits a wide range of viruses by blocking an early stage of the viral replication cycle. Cloning, characterization, and expression of Mx in vivo and in vitro have been conducted. The chicken Mx gene spans 21 kb and is made up of 14 exons and 13 introns, of which the promoter region was analyzed. The real-time PCR results showed that Mx expression was increased in chicken embryo fibroblasts (CEF) after 12- and 24-h induction with polyI: C. Induction of Mx expression by poly I: C in vivo revealed tissue-specific patterns among the chicken tissues tested. A trace expression of Mx was detected in healthy chicken liver tissues from adult chickens without inducement; the expression levels in the liver, heart, and gizzard were higher than in the muscle and kidney. This is the first report to demonstrate the expression of a glutathione-S-transferase-tagged-Mx fusion protein of 75 KDa, as well as the biological activity tested by SDS-PAGE and western blotting.

Disc and underwriting - A proposal of life underwriter in terms of insurance benefits - (디스크질환과 언더라이팅 -보장급부를 중심으로 고찰한 생명보험 언더라이터의 제안-)

  • Byun, Hye-Jin
    • The Journal of the Korean life insurance medical association
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    • v.27 no.2
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    • pp.96-106
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    • 2008
  • Herniate disc disease is one of the biggest problem in claim of insurance as well as in medical. Herniate disc disease have recently increased, and it is ranked 8th in claim reasons recently. As an underwriter and physical therapist, I want to study interrelationship of insurance and disc disease. I think it is necessary to know about knowledge of medical, so this study is given some space to structure of spine, cause of herniated disc disease, role of disc, methods of classification of disable (McBride method and AMA method), and spine disability stage. disc surgery is divided laparoscope disc surgery and spine surgery. I analysis it some factors- gender, age, occupation, re-surgery, and state of after surgery - through searching medical papers. I suggest below conclusion to underwriter because it can be useful to make questionnaire, and underwriter can expect prognosis. conclusion The negative factors of disc surgery (compare to other cases) were as follows: 1. endoscope disc surgery: $20{\sim}40year$ old man, hospitalization period more than 5 days. 2. spine surgery: $45{\sim}70year$ old woman, hospitalization period more than 15 days. 3. re-surgery experience: exist 4. working condition: a person who draws a small income, non-regular worker, working period is less than 1 year. 5. method of surgery: pedicle screw fixation. spine fusion surgery, artificial disc surgery. 6. post surgery condition: appearance of muscle weakness, paralysis, reference pain, lordosis, kyphosis, and complication. smoker or take a drink.

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Radiological and Clinical Results of Laminectomy and Posterior Stabilization for Severe Thoracolumbar Burst Fracture : Surgical Technique for One-Stage Operation

  • Kim, Myeong-Soo;Eun, Jong-Pil;Park, Jeong-Soo
    • Journal of Korean Neurosurgical Society
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    • v.50 no.3
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    • pp.224-230
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    • 2011
  • Objective : This study aimed to show the possibility of neural canal enlargement and restoration of bony fragments through laminectomy and minimal facetectomy without pediculectomy or an anterior approach, and also to prove the adequacy of posterior stabilization of vertebral deformities after thoracolumbar bursting fracture. Methods : From January 2003 to June 2009, we experienced 45 patients with thoracolumbar burst fractures. All patients enrolled were presented with either a neural canal compromise of more than 40% with a Benzel-Larson Grade of VI, or more than 30% compromise with less than a Benzel-Larson Grade of V. Most important characteristic of our surgical procedure was repositioning retropulsed bone fragments using custom-designed instruments via laminectomy and minimal facetectomy without removing the fractured bone fragments. Beneath the dural sac, these custom-designed instruments could push the retropulsed bone fragments within the neural canal after the decompression and bone fragment repositioning. Results : The mean kyphotic deformities measured preoperatively and at follow-up within 12 months were 17.7 degrees (${\pm}6.4$ degrees) and 9.6 degrees (${\pm}5.2$ degrees), respectively. The mean midsagittal diameter improved from 8.8 mm (${\pm}2.8$ mm) before surgery to 14.2 mm (${\pm}1.6$ mm) at follow-up. The mean traumatic vertebral body height before surgery was 41.3% (${\pm}12.6%$). At follow-up assessment within 12 months, this score showed a statistically significant increase to 68.3% (${\pm}12.8%$). Neurological improvement occurred in all patients. Conclusion : Though controversy exists in the treatment of severe thoracolumbar burst fracture, we achieved effective radiological and clinical results in the cases of burst fractures causing severe canal compromise and spinal deformity by using this novel custom-designed instruments, via posterior approach alone.

Experience of Single Stage Treatment of Caniosynostosis, Hypertelorism, Exophthalmos Patient (두개조기유합증, 안와격리증, 안구돌출증 환자의 동시 교정 치험례)

  • Chung, Chul Hoon;Eun, Seok Chan;Seo, Dong Kuk;Jo, Woo Sung;Park, Se Hyuck
    • Archives of Plastic Surgery
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    • v.33 no.2
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    • pp.237-240
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    • 2006
  • The simultaneous correction of the hypertelorism and exophthalmos combined with craniosynostosis is very rarely performed operative procedures in the world. The craniosynostosis is the congenital anomaly that designates premature fusion of one or more sutures in either cranial vault or cranial base. Hypertelorism is not a distinct clinical syndrome in itself, but is a physical finding secondary to facial and cranial maldevelopment and it is defined as a increase in the distance between the medial orbital walls. Exophthalmos can occur following the decrease in the size of the orbit in patients with developmental skeletal disorders such as craniofacial synostosis. The authors experienced 9-year-old male patient, who has complex cranio-facial abnormality. The craniosynostosis was oxycephaly type and primary fronto-orbital advancement surgery had been performed in other hospital. The abnormal cranial vault combined with hypertelorism and exophthalmos due to maldeveloped both orbital walls. Surgical correction was obtained by various cranio-fronto-orbital remodeling technique such as calvarial bone craniotomy, fronto-orbital advancement, paramedian resection, medial canthopexy, Tessier-Wolfe three wall orbital expansions. We achieved a quite satisfactory result both functionally and aesthetically in a complex cranio-facial deformity patient by combination and modification of previously developed various cranio-facial plasty technique and hereby report the case with brief discussion and review of literature.