• 제목/요약/키워드: prolonged culture

검색결과 164건 처리시간 0.023초

Glyphosate 독성(毒性): II. corydalis Sempervirens와 토마토의 세포배양체(細胞培養體)에서 EPSP-synthase의 활성(活性)에 미치는 영향(影響) (Glyphosate Toxicity: II. EPSP-synthase Activity in Cell Suspension Culture of Corydalis Sempervirens and Lycopersicon Esculentum)

  • 김태완;니콜라스 암라인
    • 한국잡초학회지
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    • 제15권2호
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    • pp.148-153
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    • 1995
  • Glyphosate (N-[phosphonomethyl]glycine)를 토마토(Lycopersicon esculentum Mil)의 동화부위(同化部位)에 부분처리(部分處理)하거나 전(全) 식물체(植物體)에 분무처리(噴霧處理)하였을 때 EPSP-synthase의 활성(活性) 감소(減少)가 나타났다. EPSP-synthase의 활성(活性)은 처리된 식물체(植物體)의 엽록소(葉綠素)의 감소보다 시기적으로 먼저 나타나는 현상이었다. EPSP-synthase의 활성(活性)은 glyphosate처리(處理)에 민감한 효소(酵素)로서 토마토의 세포현탁배양조직(細胞顯濁培養組織)과 분열조직(分裂組織)간에는 활성(活性)의 차이가 없없다. EPSP-synthase의 활성은 4-6 nkat/mg protein 정도이었다. EPSP-synthase의 활성(活性)억제는 glyphosate 처리(處理) 36시간 후 부터 나타나기 시작하였고, 엽록소(葉綠素)의 감소는 처리 48 시간 후 부터 나타나기 시작하였다. 세포현탁배양(細胞顯濁培養)에서 치사농도(致死濃度) 이하(以下)에서 glyphosate는 생체중(生體重)을 저하(低下)시켰으며 생육단계(生育段階) 중 lag-phase를 연장(延長)시켜 생육(生育)이 더디도록 하였다. Glyphosate 존재(存在)하에서 생체중(生體重)은 계대(繼代)배양 후 14일이 지난 뒤에 생체중(生體重)이 최고(最高)에 달하였다. EPSP-synthase에 대한 gly-phosate의 억제(抑制)효과는 lag-phase에서 심하게 나타났다.

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자유생활아메바 Naegleria fowleri의 계대감염에 의한 병원성의 변화에 관한 연구 (Changes in the pathogenicity of Naegleria fowleri by serial brain passage in mice)

  • 이득기;임경일
    • Parasites, Hosts and Diseases
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    • 제21권2호
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    • pp.234-240
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    • 1983
  • 장기간 시험관내에서 무균적으로 배양하여 병원성이 약화된 자유생활아메바 naegleria fowleri를 마우스에 연속적으로 감염시켜 그 병원성의 증감여부를 관찰하고자 하였다. 체증 18∼22gm의 백색 마우스를 secobarbital로 마취시키고 오른쪽 비강에 Naegleria fowleri를 떨어뜨려 감염시켰다. 시험관내에서 7년이상 CGVS배지에서 계대배양된 0주와 마우스에 감염시켜 뇌조직을 2번 통과시킨 2-1주의 병원성을 비교하였다. 행동둔화, 자극에 민감, 회전운동, 사지마비등 여ㅓ 증상이 나타났고, 뇌의 오른쪽 앞쪽 부위에 심한 염증 및 괴사를 발견하였다. 이러한 증상과 병소는 0주 감염군보다 2-1감염군에서 보다 빨리, 보다 심하게 발생되었음을 관찰하였다. 감염 7일 후부터 감염된 마우스가 사망하기 시작하였으며 2-1주 감염군에서 0주 감염군 보다 생존기간이 짧았으며 감염 13일 후부터 사망한 예에 있어서는 육안적으로 뇌에서 병소가 없었고 폐장에서 염증이 심하였음을 관찰하였다. 마우스에 비강을 Naegleria fowleri를 감염시켜 전형적인 원발성 아메바성 뇌수막염을 발생시킬 수 있었다. 장기간 시험관에서 무균적으로 계대배양된 Naegleria fowleri는 병원성이 약화되어 있었고 이 아메바를 마우스에 연속적으로 감염시켜 뇌조직을 통과시킴으로 다시 병원성이 증강됨을 관찰하였다.

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글루타르알데하이드 고정 소심 낭막에서의 내피세포 증식에 대한 글루탕산 및 파라벤용액의 효과 (Effect of L-Glutamic Acid and Paraben Solution on the Endothelial Cell Proliferation in the Glutaraldehyde- Fixed Bovine Pericardium)

  • 김범식;이문환;유세영;김원곤
    • Journal of Chest Surgery
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    • 제29권1호
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    • pp.7-13
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    • 1996
  • The conventional glutaraldehyde (GA) fixation method of tissue valves is considered to be responsible for accelerated valve degeneration. The release of toxic GA from the valve tissue is believed to limit endothelial cell (EC) ingrowth. Removal of toxic GA by reaction with L-glutamic acid and storage in a Paraben solution may offer good EC growth. To investigate the conditions for endothelialization of tissue valves, the growth properties of ECs on the conventionally and alternatively treated pericardial tissue were compared. Conventional preparation included zero-pressure fixation for 72 hours in phosphated-buffered saline (PBS) solution containing 0.5% GA at 4$^{\circ}C$ and storage into PBS containing 0.2% GA(group I). Alternatively treated pericardial tissues were divided into three postfixation treatment groups : (1) storage in PBS solution containing Paraben(group II), (2) treatment with PBS containing 8$^{\circ}C$ L-glutamic acid(PH 7.35) and storage in PBS solution containing Paraben (g oup III), (3) treatment with L-glutamic acid dissolved in distilled water (PH 3.5) (group IV). Pericardial tissue were transferred into the 24-well plate after storage for 4 weeks. ECs were harvested enzymatically from the bovine pulmonary artery and grown to confluence on culture flask surfaces. Detached ECs by trypsin were incubated into the each well of the 24-well plate including test pericardial tissues. Cells were detached by trypsin, 1, 2, 3, 5, 7 days after incubation and counted on the hemacytometer. Cell viability test was performed by frypan-blue exclusion method. Acute cell death in the group I were found even after prolonged washing. The group II showed prolonged cell survival compared with the group I. Both group III and group IV showed better cell growth than group II. There was no statistically significant difference between group III and group IV method in terms of EC growth. This results suggest that treatment by L-glutamic ac id and storage in a Paraben solution be a promising approach for improvement of durability of GA-treated tissue valves.

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소아 요로 감염 및 의심 환아에서 신 실질 병변 및 방광요관 역류와 임상 변수와의 연관성 (The relationships between clinical variables and renal parenchymal disease in pediatric clinically suspected urinary tract infection)

  • 변정림;이상택;정소정;김교순
    • Clinical and Experimental Pediatrics
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    • 제53권2호
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    • pp.222-227
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    • 2010
  • 목 적 : 본 연구는 소아 요로 감염증에서 치료 전 발열 기간과 치료 후 발열 기간 등의 임상변수와 혈액 및 소변 등의 검사 결과가 신 실질 병변 및 방광요관 역류 등을 예측하는 데 있어서 인자로 작용할 수 있는지를 평가해 보고자 하였다. 방 법 : 2005년 7월부터 2008년 7월까지 첫 번째 열성 요로 감염으로 본원 소아과에 입원한 1개월부터 만 17세까지의 환아 180명을 대상으로 하였다. 환아의 혈액 검사 소견 중 C-반응 단백, 백혈구수와 소변 검사 중 배양 검사 결과, 소변 질산염 및 치료전 발열기간, 치료 후 발열 기간 등을 변수로 하여 신장 초음파 배뇨 방광, 요도 조영술, 신 스캔 등의 영상 검사 결과와 비교하여 연관성을 평가하였다. 결 과 : C-반응 단백 수치가 높고 백혈구 증가증이 있으며 입원 치료 후 긴 발열 기간을 가진 경우 신 실질 병변이 유의하게 증가하였고 3단계 이상의 방광 요관 역류 발생도 높았다. 신 스캔 검사상 이상 소견을 보인 경우는 신 스캔 검사상 정상 소견을 가진 군과 비교하였을 때 3단계 이상의 방광 요관 역류가 더 증가되어 있었다. 결 론 : C-반응 단백 수치, 백혈구 증가증, 입원 치료 후 긴 발열기간 등의 임상 변수는 신 실질 병변과 3단계 이상의 방광 요관 역류의 예측 인자가 될 수 있을 것으로 사료된다.

양수 세포를 이용한 인간배아줄기세포의 배양 (Human Amniotic Fluid Cells Support Expansion Culture of Human Embryonic Stem Cells)

  • 김희선;설혜원;안희진;오선경;구승엽;김석현;최영민;김정구;문신용
    • Clinical and Experimental Reproductive Medicine
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    • 제31권4호
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    • pp.261-271
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    • 2004
  • Objective: This study was performed to evaluate the possibility of prolonged culture of human embryonic stem cells (hESC; SNUhES2) on human amniotic fluid cells (hAFC), which had been storaged after karyotyping. Method: The hAFC was prepared for feeder layer in the presence of Chang's medium and STO medium (90% DMEM, 10% FBS) at $37^{circ}C$ in a 5% $CO_2$ in air atmosphere. Prior to use as a feeder layer, hAFC was mitotically inactivated by mitomycin C. The hESCs on hAFC were passaged mechanically every seven days with ES culture medium (80% DMEM/F12, 20% SR, bFGF). Results: The hAFC feeder layer support the growth of undifferentiated state of SNUhES2 for at least 59 passages thus far. SNUhES2 colonies on hAFC feeder appeared slightly angular and flatter shape as compared with circular and thicker colonies observed with STO feeder layer and showed higher level with complete undifferentiation in seven days. Like hESC cultured on STO feeders, SNUhES2 grown on hAFC expressed normal karyotype, positive for alkaline phosphatase activity, high telomerase activity, Oct-4, SSEA-3, SSEA-4, Tra-1-60 and Tra-1-81 and formed embryoid bodies (EBs). Conclusion: The hAFC supports undifferentiated growth of hESC. Therefore, these results may help to provide a clinically practicable method for expansion of hESC for cell therapies.

메기(Silurus asotus)에 투여한 멜라민의 체내함량 변화 (Accumulation and Depletion of Melamine Through Experimental Feeding in Catfish Silurus asotus)

  • 김풍호;조미라;이희정;김경덕;하광수;유현덕;유홍식;이두석;윤호동
    • 한국수산과학회지
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    • 제44권6호
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    • pp.577-583
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    • 2011
  • In 2007, pet food contaminated with melamine caused hundreds of dogs and cats to develop renal failure all over the world. In 2008, over 294,000 infants consumed infant formula and developed kidney stones in China. Further investigation revealed that fish feed also contained melamine; this raised concerns about melamine residues in edible fish tissues, which could have caused the urinary tract stone epidemic. In Korea, catfish fed with assorted feed that included cuttlefish organs that contained melamine developed whitening syndrome and fell dead in some populations in 2008. This event raised suspicions about the toxicity of melamine and all feeds containing melamine were immediately recalled. In this study, we investigated the rates of melamine accumulation and depletion in muscle and viscera of catfish to propose proper withdrawal periods. One group of catfish was fed a commercially available diet that contained 30, 100 and 300 mg melamine per kg diet for 14 days. To investigate residual melamine contents in muscle and viscera, other experimental groups were fed a melamine free diet after being fed melamine for 7 days. The residual amount of melamine was analyzed by LC-MS/MS. The melamine concentration in muscle was estimated to be 3.7 mg/kg after 6 days of feeding with a diet containing 300 mg melamine/kg. After 2 days of culture with a melamine free diet, the residual melamine was depleted and the concentration had decreased from 1.15 mg/kg to 0.19 mg/kg in the muscle of catfish fed a diet containing 300 mg melamine/kg for 7 days. The residual amount of melamine was reduced to 0.03 mg/kg in muscle after 7 days of culture with a melamine free diet and was undetectable after a prolonged culture period of 14 days. Catfish tend to excrete melamine rapidly after oral administration and changes in body color were not observed during the short dosing period.

돼지난자의 체외성숙과 수정에 있어서 Xanthine-Xanthine Oxidase System의 이용 (Use of a Xanthine-Xanthine Oxidase System on in vitro Maturation and Fertilization in the Pig)

  • Sa, S. J;H. T. Cheong;Lee, S. Y.;Lee, J. H.;I. S. Ryu;B. K. Yang;Kim, C. I.;Park, C. K.
    • 한국가축번식학회지
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    • 제27권1호
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    • pp.77-85
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    • 2003
  • 본 연구는 xanthine(X)-xanthine oxidase(XO) system하에서 돼지 난자의 체외성숙과 체외수정에 대한 catalase의 영향을 검토하였다. 그 결과 돼지 난포난자가 X 또는 XO하에서 배양되었을 때, 난포난자의 성숙율은 다른 배양시간에도 불구하고 catalase 첨가 유무에 따른 유의적인 차이를 나타내지 않았다. 그렇지만, X-XO-catalase system하에서 배양한 경우 유의적으로 높은 성숙율을 얻었다(P<0.05). 퇴행난자의 비율은 배양기간이 늘어남에 따라 증가되었으며, 배양 120시간에서는 catalase 첨가시보다 무첨가시에 유의적으로 높았다. 다른 한편으로, 단위발생 난자들이 배양 72시간에 높은 비율로 관찰되었지만, 다양한 배양시간에서 catalase 첨가유무에 따른 차이는 발견되지 않았다. 또 다른 실험에서, 동결-응해된 돼지 정자가 체외수정을 위해 X-XO system으로 처리되었다. 난자투명대에 대한 정자침입율은 none (P<0.05), XO, X+XO하에서 체외수정시 catalase 무첨가시보다 첨가시에 높게 나타났다. 다른 한편으로, 돼지정자가 none, X, XO, X+XO로 처리되었을 때, lipid peroxidation은 catalase 첨가시보다 무첨가시에 높은 비율로 나타났으며, 그 결과 정자침입과 lipid peroxidation에서의 변화가 상반되는 양상을 보였다. 그렇지만, 모든 조건하에서 정자의 sulfhydry (-SH) group의 함량은 catalase 첨가시에 높게 측정되었다. 난자의 투명대에 대한 정자의 접착 정도는 salt-stored 돼지 난자에 대한 정자접착을 통해서 평가되었으며, control group의 경우 X, XO, X+XO group에 비해 높은 정자접착율이 관찰되었다. 그렇지만, catalase 첨가유무에 따른 유의적인 차이는 인정되지 않았다. 본 연구의 결과는 X-XO-catalase system에 대한 난포난자와 정자의 노출이 돼지에서의 체외성숙과 체외수정을 촉진시키는 것으로 생각된다.

Genetic Analysis of Spontaneous Lactose-Utilizing Mutants from Vibrio vulnificus

  • Baek, Chang-Ho;Lee, Ko-Eun;Park, Dae-Kyun;Choi, Sang-Ho;Kim, Kun-Soo
    • Journal of Microbiology and Biotechnology
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    • 제17권12호
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    • pp.2046-2055
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    • 2007
  • Wild-type V. vulnificus cannot grow using lactose as the sole carbon source or take up the sugar. However, prolonged culture of this species in media containing lactose as the sole carbon source leads to the generation of a spontaneous lactose-utilizing (LU) mutant. This mutant showed strong ${\beta}$-galactosidase activity, whereas the wild-type strain showed a barely detectable level of the activity. A mutant with a lesion in a gene homologous to the lacZ of E. coli in the bacterium no longer showed ${\beta}$-galactosidase activity or generated spontaneous LU mutants, suggesting that the lacZ homolog is responsible for the catabolism of lactose, but the expression of the gene and genes for transport of lactose is tightly regulated. Genetic analysis of spontaneous LU mutants showed that all the mutations occur in a lacI homolog, which is located downstream to the lacZ and putative ABC-type lac permease genes. Consistent with this, a genomic library clone containing the lad gene, when present in trans, made the spontaneous LU mutants no longer able to utilize lactose as the sole carbon source. Taken together with the observation that excessive amounts of exogenously supplemented possible catabolic products of lactose have negative effects on the growth and survivability of V. vulnificus, we suggest that V. vulnificus has evolved to carry a repressor that tightly regulates the expression of lacZ to keep the intracellular toxic catabolic intermediates at a sublethal level.

Genistein from Vigna angularis Extends Lifespan in Caenorhabditis elegans

  • Lee, Eun Byeol;Ahn, Dalrae;Kim, Ban Ji;Lee, So Yeon;Seo, Hyun Won;Cha, Youn-Soo;Jeon, Hoon;Eun, Jae Soon;Cha, Dong Seok;Kim, Dae Keun
    • Biomolecules & Therapeutics
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    • 제23권1호
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    • pp.77-83
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    • 2015
  • The seed of Vigna angularis has long been cultivated as a food or a folk medicine in East Asia. Genistein (4',5,7-trihydroxyisoflavone), a dietary phytoestrogen present in this plant, has been known to possess various biological properties. In this study, we investigated the possible lifespan-extending effects of genistein using Caenorhabditis elegans model system. We found that the lifespan of nematode was significantly prolonged in the presence of genistein under normal culture condition. In addition, genistein elevated the survival rate of nematode against stressful environment including heat and oxidative conditions. Further studies demonstrated that genistein-mediated increased stress tolerance of nematode could be attributed to enhanced expressions of stress resistance proteins such as superoxide dismutase (SOD-3) and heat shock protein (HSP-16.2). Moreover, we failed to find genistein-induced significant change in aging-related factors including reproduction, food intake, and growth, indicating genistein exerts longevity activity independent of affecting these factors. Genistein treatment also led to an up-regulation of locomotory ability of aged nematode, suggesting genistein affects healthspan as well as lifespan of nematode. Our results represent that genistein has beneficial effects on the lifespan of C. elegans under both of normal and stress condition via elevating expressions of stress resistance proteins.

포스트 코로나19 언택트 시대 대응을 위한 AI 챗봇 구축방법에 관한 연구 (A Study on the Method of Implementing an AI Chatbot to Respond to the POST COVID-19 Untact Era)

  • 정천수;정지환
    • 한국IT서비스학회지
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    • 제19권4호
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    • pp.31-47
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    • 2020
  • Recently, as the COVID-19 has spread and prolonged worldwide, the 'Untact' society is becoming routinized, and various smart technologies are leading to the spread of the 'Ontact' culture. This is because the desire of consumers to purchase a product and use the service has increased while minimizing the direct contact. In order to quickly respond to this circumstance, the percentage of the companies which are adopting Chatbot in various fields such as orders, delivery, and inquiries is increasing and they are getting a positive result. However as the demand for building Chatbot increases dramatically, there are many confusions among the companies which want to introduce Chatbot to their system, due to the lack of professional technicians and difficulties in understanding AI technologies and how to build them effectively. I believe that in the post COVID-19 era, much more companies will adopt Chatbot, and this will intensify the problem. The purpose of this study was to derive the needs for a guide on the method of buiilding a Chatbot through considering the prior research on Chatbot and analysis of the recent surge in the use of Chatbot services related to COVID-19. There are implications to presenting 5 phases of universal Chatbot implementation methodology using the platform to the stakeholders who want to introduce Chatbot to their customer so that they can understand and build Chatbot more easily and use AI Chatbot actively in response to the POST COVID-19 era.