• Title/Summary/Keyword: Aminopurine

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Inhibition of ER Stress by 2-Aminopurine Treatment Modulates Cardiomyopathy in a Murine Chronic Chagas Disease Model

  • Ayyappan, Janeesh Plakkal;lizardo, Kezia;Wang, Sean;Yurkow, Edward;Nagajyothi, Jyothi F
    • Biomolecules & Therapeutics
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    • v.27 no.4
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    • pp.386-394
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    • 2019
  • Trypanosoma cruzi infection results in debilitating cardiomyopathy, which is a major cause of mortality and morbidity in the endemic regions of Chagas disease (CD). The pathogenesis of Chagasic cardiomyopathy (CCM) has been intensely studied as a chronic inflammatory disease until recent observations reporting the role of cardio-metabolic dysfunctions. In particular, we demonstrated accumulation of lipid droplets and impaired cardiac lipid metabolism in the hearts of cardiomyopathic mice and patients, and their association with impaired mitochondrial functions and endoplasmic reticulum (ER) stress in CD mice. In the present study, we examined whether treating infected mice with an ER stress inhibitor can modify the pathogenesis of cardiomyopathy during chronic stages of infection. T. cruzi infected mice were treated with an ER stress inhibitor 2-Aminopurine (2AP) during the indeterminate stage and evaluated for cardiac pathophysiology during the subsequent chronic stage. Our study demonstrates that inhibition of ER stress improves cardiac pathology caused by T. cruzi infection by reducing ER stress and downstream signaling of phosphorylated eukaryotic initiation factor ($P-elF2{\alpha}$) in the hearts of chronically infected mice. Importantly, cardiac ultrasound imaging showed amelioration of ventricular enlargement, suggesting that inhibition of ER stress may be a valuable strategy to combat the progression of cardiomyopathy in Chagas patients.

Embryo Culture of Taxus wallichiana (Zucc.)

  • Datta Mukul Manjari;Jha Sumita
    • Journal of Plant Biotechnology
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    • v.6 no.4
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    • pp.213-219
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    • 2004
  • Zygotic embryos were excised from immature and mature seeds of the Himalayan yew, Taxus wallichiana. The embryos germinated precociously when kept in darkness for 5 weeks and developed into full seedlings within 10-12 weeks. The highest rate of embryo germination ($81\%$) was obtained in modified Lloyd & McCown' s woody plant medium containing macro and micronutrients at half strength supplemented with $1\%$ activated charcoal, which supported both the best embryonic growth ($43\%$) and seedling development ($32\%$). However, the supplementation of basal media with kinetin, thidiazuron, 6-benzyl aminopurine or $GA_3$ had no effect on the germination of the embryos. The embryos derived from immature seeds germinated but the frequency of embryonic growth was better in mature seeds. Stratification of seeds effected precocious germination of embryos. Seeds kept at $4^{\circ}C$ for 1 week germinated earlier and at a higher frequency irrespective of the stage of seed maturity, while the germination rate declined with prolonged cold treatment for 1 month at that same temperature. Analysis of taxanes in germinating seedlings revealed that root tissues contained high levels of taxol, 10-deacetyl-baccatin ill and baccatin ill as compared to shoots. Thus embryo culture technique appears to overcome the lengthy dormancy requirement of T. wallichiana seeds.

Plant Regeneration from the Stem Tissue of Orostachys japonicus A. Berger (바위솔의 줄기조직으로부터 식물체 재분화)

  • 최상욱;남상해;양기종;조무제;양민석
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.2
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    • pp.65-68
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    • 1994
  • Plant regeneration from the stem tissue of Orostachys japonicus A. Beiger was investigated. The calli derived from shoot apex when apex when cultured on Murashige and Skoog (MS) medium supplemented with 4mg/L 2,4-dichlorophenoxyacetic acid (2,4-D)and 2 mg/L benzyl aminopurine (BAP). The calli were developed into shoot to MS medium with 0.5mg/L NAA and 2mg/L and into root with 1mg/L kinetin. The reddish pigment which might be essential for the rootregeneration was observed in the tip of regenerated root.

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Effect of Activation Method and Culture Medium on the Development of Porcine Nuclear Transfer Embryo using Fetal Fibroblast

  • Im, Gi-Sun;Yang, Byoung-Chul;Park, Jin-Ki;Kim, Hyun-Ju;Chang, Won-Kyung;R. S. Prather;B. N. Day
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.66-66
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    • 2001
  • Since the first birth of pig derived from embryonic cells by nuclear transfer, many researches to produce cloned pig have been carried out. Recently, two reports about the birth of somatic cell cloned pigs using in vivo oocytes and also Betthauser et al. (2000) reported the birth of somatic cell cloned pigs using in vitro oocytes. So here we investigated the effect of activation method and culture medium on in vitro development of porcine nuclear transfer embryo using fetal fibroblast. Oocytes derived from slaughter house obtained ovaries were matured for 42 to 44 h in TCM 199. Matured oocytes were denuded using 0.1% hyaluronidase and then Oocytes with the first polar body were used for enucleation by aspirating the first polar body and adjacent cytoplasm in TCM 199 supplemented with 7.5 $\mu\textrm{g}$ cytochalasin B. Petal fibroblast cells were prepared from 35 days old fetus. To be used as donor cells, fetal fibroblast cells were serum starved for 3 to 5 days and then isolated into single co:1 by trypsinization. Nuclear transfer embryos were fused using 2 times 1.25㎸ for 30$mutextrm{s}$. Fused NT embryos were activated with calcium ionophore (CI) and 6-dimethyl-aminopurine (6-DMAP). Activated oocytes were cultured in NCSU 23 or BECM 3 for 6 days. There was no significant difference between chemical activation and no chemical activation for blastocyst development rate(11.6 vs. 14.8%). However, cell number was significantly higher when NT embryos were activated with CI and 6-DMAP (31.2 vs. 22.6). When NT embryos were cultured in NCSU 23 or BECM 3, blastocyst development rate was 16.4 and 13.2%, respectively, and cell number was 31.5 and 24.1, respectively. These results suggest that chemical activation after fusion and culture in NCSU 23 could increase cell number of porcine NT embryos.

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칡소 귀세포를 이용한 핵이식란의 배양방법이 배반포 발달율과 수태율에 미치는 영향

  • 윤종택;이호준;최은주
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.65-65
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    • 2001
  • 본 연구는 칡소 귀세포를 공여핵으로 이용한 체세포 복제송아지 생산에 있어서 배양방법이 배발생 및 배반포 발달율에 미치는 영향과 체세포 복제란의 이식후 수태율에 미치는 영향을조사하여 복제송아지의 생산 효율을 제고하고자 실시하였다. 실험에 공시된 공여핵은 칡소 의 귀세포를 회수하여 10%FBS가 첨가된 DMEM배지에서 3-4일 동안 배양하여 monolayar Confluent 형성 후 0.25% trypsin을 처리하여 준비하였으며 공여세포는 적어도 passage가 5회 이상의 세포만을 사용하였다. 복제수정란의 생산은 18-20시간 동안 체외성숙 된 난자의 핵을 제거하고 공여핵을 주입하여 2.2kv/cm, 10$\mu\textrm{s}$의 전압으로 2회 자극함으로 융합하였으며, 융합된 난자는 5$\mu\textrm{g}$/$m\ell$ ionomycin에서 4분간, 1.9mM 6-dimethyl aminopurine에서 4시간동안 배양하여 활성화처리를 하였다. 핵이식수정란의 배양은 39$^{\circ}C$, 5%$CO_2$ incubator에서 처리구 I은 CRlaa에서 4일간 배양 후 CR2aa배지에서 배양, 처리구II는 CRlaa에 4일간 배양후 CR2aa배지에 cumulus cell과 공배양, 처리구III은 CR2aa 배지에 camulus cell과 함께 배양하였다. 수정란이식은 발정발현 7일째에 비외과적 방법으로 젖소 미경산우에 이식하였으며 이식란수는 2~4개의 핵이식된 수정란을 이식하였다. 임신진단은 45~60일 사이에 직장검사 및 초음파 진단기를 이용하여 실시하였다. 배양방법에 따른 배발생율은 처리구 I에서 92.2 %(83/90)으로 처리구II와 III의 62.4%(63/101)와 77.8%(144/185)에 비하여 높게 나타났으나 배반포 발달율은 처리구II와III에서 65.1%(41/63)와 50.0%(72/144)로 처리구 I의 30.1%(25/83)보다 높게 나타났다. 각 처리구에 따른 수정란 이식후 수태율은 처리구II와 III에서 공히 20%의 수태율을 나타낸 반면 처리구 I에서는 수태가 되지 않았다. 따라서 체세포 복제수정란의 생산에 있어서 배반포 발달율과 수태율을 높이기 위해서는 단순배양보다 공배양이 더 효과적인 것으로 사료되지만 이런 결과가 복제송아지 생산효율에 있어서도 효과적일지는 향후 더 많은 연구가 있어야 할 것으로 사료된다.

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Plant Regeneration from Embryogenic Callus of Miscanthus spp. (억새(Miscanthus spp.) 배발생 캘러스로부터 식물체 재분화)

  • Kim, Kwang-Soo;Kwon, Da-Eun;Lee, Ji-Eun;Cha, Young-Lok;Moon, Youn-Ho;Kang, Yong-Ku
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.89-89
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    • 2018
  • 우리나라, 중국 및 일본을 포함한 동북아시아가 원산이며 바이오매스량이 많은 억새(Miscanthus spp.)는 바이오에너지 생산을 위한 원료작물로서 가치가 높아, 바이오에탄올 생산용 원료작물로 주목을 받고 있다. 독일 등 유럽과 미국에서는 바이오에탄올 생산용 작물로 주로 종간 교잡 이질 3배체인 불임성 억새(M. x giganteous)를 대상으로 연구하고 있다. 이렇게 단일유전형을 갖는 품종의 재배에는 특정 병과 해충에 약하며 자연재해에도 취약성을 나타내므로 억새가 바이오에너지작물로 자리 잡기 위해서는 다양한 유전형의 억새 품종 개발이 필요하다. 본 연구는 우리나라의 자생 억새 3종을 기내배양하고 탈분화 및 재분화 시스템을 구축하여 억새 품종 육성 시 효율을 높이기 위해서 실시하였다. 억새 종자로부터 캘러스의 유도는 MS배지보다 N6배지 에서 좋았으며, 식물생장조절제로 2,4-Dichlorophenoxyacetic acid (2,4-D)와 6-Benzyl aminopurine (BA)를 조합처리한 처리구보다 2,4-D만을 단독처리하였을 때 캘러스 유도율이 더 높았다. 억새 종에 따른 캘러스 유도율은 물억새가 가장 낮고, 거대억새가 가장 높았으며, 3 ~ 5 mg/L의 2,4-D가 첨가된 N6배지에서 배발생 캘러스(embryogenic callus)가 발생하였다. 억새 신초 및 줄기의 절간에서의 캘러스 유도율은 전반적으로 종자에 비하여 낮았으며, 미성숙화기로부터의 캘러스 유도는 억새 종에 따른 차이가 없었으며, 5mg/L의 2,4-D가 첨가된 배지에서 캘러스 유도율이 가장 높게(90 ~ 95%) 나타났다. 형성된 배발생 캘러스로부터 식물체의 재분화는 N6배지에서는 재분화 식물체가 발생하지 않았고, 1 ~ 3mg/L의 BA와 0.1ml/L의 1-Naphthaleneacetic acid (NAA)가 첨가된 MS배지에서만 식물체가 재분화되였다.

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Histological Characteristics of Somatic Embryos in Melon (Cucumis melo L.) (멜론 체세포배의 조직학적 특징)

  • Choi, Pil Son;Kwon, Suk Yoon
    • Korean Journal of Plant Resources
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    • v.26 no.4
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    • pp.511-515
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    • 2013
  • Hypocotyls explants of melon seedling were cultured on Murashige and Skoog's (MS) medium supplemented with 1 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.5 mg/L benzyl aminopurine (BA) for 6 weeks to produce somatic embryos. In somatic embryos produced through intervening bright yellow friable (BYF) from the explants, somatic embryos with two-cotyledon (26%) and horn-type cotyledon (74%) were observed. The procambial strand of cotyledons was originated from circular procambial tissues of lower hypocotyls. The circular procambial independently divided into two procambial strand at the edge of cotyledonary-node, and then connected to each cotyledon to form somatic embryos with two-cotyledon. When cotyledon was horn-type, the circular procambial strand in lower hypocotyls would continuously remain connected to the cotyledon. However, somatic embryos with two or horn type cotyledon formed an abnormal shoot apex without the tunica-corpus structure or dome shape in the inter-cotyledonary area. These results demonstrated that the variation of cotyledon in somatic embryos was closely related to procambial tissue differentiation and shoot apical formation.

Plant Regeneration of Iris koreana Nakai through Organogenesis for Ex-situ Conservation

  • Bae, Kee-Hwa;Yun, I-Seul;Jung, Ji-Sun;Kim, Chan-Beom;Kim, Hye-Won;Hong, Yong-Sik;Oak, Min-Kyeong;Kim, Hak-Koo;Lee, Ju-Hui
    • Journal of Forest and Environmental Science
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    • v.37 no.4
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    • pp.304-308
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    • 2021
  • Iris koreana (Iridaceae) is an endangered plant native to Korea. In order to develop an in vitro propagation method, we investigated the effect of 2,4-dichlorophenoxy acetic acid (2,4-D) and a-naphthalene acetic acid (NAA) on callus induction in different I. koreana tissues. In addition, we also investigated the effect of 2,4-D and Benzyl aminopurine (BA) treatments on adventitious shoot induction in viable calli and the effect of indole-3-butyric acid (IBA) on root formation in viable shoots. We found that callus production was highest with 1.0 mg/L NAA (94.4% cultured rhizome explants), and adding low concentrations of 2,4-D to BA containing media significantly increased the frequency of shoot primordial formation. The best rooting results were obtained with 1.0 mg/L IBA, on which 98% of regenerated shoots developed roots and produced an average of 7.4 roots within 45 days. This in vitro propagation protocol will be useful for conservation, as well as for mass propagation.

Influence of Medium and Plant Growth Regulator on Micropropagation Efficiency in Blueberry (블루베리의 미세번식에서 배지와 식물생장조절제의 영향)

  • Kim, Hwa Young;Kang, Sun Pil;Hong, Sae Jin;Eum, Hyang Lan
    • Journal of Bio-Environment Control
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    • v.24 no.3
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    • pp.167-172
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    • 2015
  • The aim of this study was to develop an effective production system of blueberry plants by using tissue culture technique. Murashige and skoog medium (MS) and woody plant medium (WPM) were compared for shoot formation of highbush blueberries. Also medium supplemented with zeatin/2-isopentenyl adenine (2iP)/benzyl aminopurine (BA) (1, 2/10, 15/4, $6mg{\cdot}L^{-1}$)and zeatin/2iP/BA (0.5/10, 15/$0.05mg{\cdot}L^{-1}$) as plant growth regulators to determine the effect of shoot formation and shoot proliferation, respectively. The shoot explants cultured on WPM showed higher shoot formation rates, more number of nodes, and longer root length than those on MS medium during the primary culture. Shoots were not formed when the explants were cultured on the medium without plant growth regulators or on only BA. The shoot explants cultured on the medium supplemented with 2iP showed low rates of shoot formation. On the other hand, zeatin was the most effective for shoot formation and growth of the explants. Also influence of different cytokinins (zeatin, 2iP) on the shoot proliferation of subcultured shoot explants was studied. There was no significant difference among the different concentrations of zeatin in the rate of shoot formation and number of shoots. However at higher concentration of zeatin, number of nodes was increased, and shoot length was shorted. The proper concentrations of zeatin for shoot propagation in subculture were found to be $0.5mg{\cdot}L^{-1}$ and $1mg{\cdot}L^{-1}$.