• 제목/요약/키워드: Replication protein A

검색결과 327건 처리시간 0.032초

Plasma Surface Modification of Patterned Polyurethane Acrylate (PUA) Film for Biomedical Applications

  • Yun, Young-Shik;Kang, Eun-Hye;Yun, In-Sik;Kim, Yong-Oock;Yeo, Jong-Souk
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2015년도 제49회 하계 정기학술대회 초록집
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    • pp.223.2-223.2
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    • 2015
  • Polyurethane acrylate (PUA) has been introduced to utilize as a mold material for sub-100 nm lithography as it provides advantages of stiffness for nanostructure formation, short curing time, flexibility for large area replication and transparency for relevant biomedical applications. Due to the ability to fabricate nanostructures on PUA, there have been many efforts to mimic extracellular matrix (ECM) using PUA especially in a field of tissue engineering. It has been demonstrated that PUA is useful for investigating the nanoscale-topographical effects on cell behavior in vitro such as cell attachment, spreading on a substrate, proliferation, and stem cell fate with various types of nanostructures. In this study, we have conducted surface modification of PUA films with micro/nanostructures on their surfaces using plasma treatment. In general, it is widely known that the plasma treated surface increases cell attachment as well as adsorption of ECM materials such as fibronectin, collagen and gelatin. Effect of plasma treatment on PUA especially with surface of micro/nanostructures needs to be understood further for its biomedical applications. We have evaluated the modified PUA film as a culture platform using adipose derived stem cells. Then, the behavior of stem cells and the level of adsorbed protein have been analyzed.

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원핵세포에서 신호물질 및 조절인자로서의 3',5'-Cyclic Adenosine Monophosphate의 역할 (3',5'-Cyclic Adenosine Monophosphate (cAMP) as a Signal and a Regulatory Compound in Bacterial Cells)

  • 천세진;석영재;이규호
    • 한국미생물·생명공학회지
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    • 제34권4호
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    • pp.289-298
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    • 2006
  • 3',5'-cyclic adenosine monophosphate (cAMP) is an important molecule, which mediates diverse cellular processes. For example, it is involved in regulation of sugar uptake/catabolism, DNA replication, cell division, and motility in various acterial species. In addition, cAMP is one of the critical regulators for syntheses of virulence factors in many pathogenic bacteria. It is believed that cAMP acts as a signal for environmental changes as well as a regulatory factor for gene expressions. Therefore, intracellular concentration of cAMP is finely modulated by according to its rates of synthesis (by adenylate cyclase), excretion, and degradation (by cAMP phosphodiesterase). In the present review, we discuss the bacterial physiological characteristics governed by CAMP and the molecular mechanisms for gene regulation by cAMP. Furthermore, the effect of cAMP on phosphotransferase system is addressed.

습사료와 부상사료에 대한 강도다리(Platichthys stellatus)의 성장과 소화 특성 (Growth Performance and Digestive Characteristics of Starry Flounder Platichthys stellatus on the Moist and Extruded Pellets)

  • 김병기
    • 한국수산과학회지
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    • 제45권6호
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    • pp.679-685
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    • 2012
  • A study was carried out to observe the effects of feed types on the growth, feed preference, and enteric feed transition rate of juvenile starry flounder, Platichthys stellatus for 45 days. Fifty fish (avg. 135 g) were stocked each in replication, and fed a commercial extruded pellet diet (EP, 45% protein) and a moist pellet diet (MP, 65% raw mackerel+35% feed powder in wet basis), respectively. The MP presented the higher performance than that of the EP on the feed efficiency ($68.3{\pm}0.9%$ for EP and $92.3{\pm}4.3%$ for MP) and the specific growth rate ($1.07{\pm}0.07$ for the EP and $1.20{\pm}0.05%$ for the MP). In contrast, the EP showed the higher feed preference in terms of the daily feed intake ($1.57{\pm}0.08%$ for the EP and $1.30{\pm}0.01$ for the MP) and the ad libitum feeding rate after a fast of 72 hours (1.73% for the EP and 1.35% for the MP). The feed transition rate through intestinal canals decreased exponentially in both the EP and the MP, showing the faster transition rate with the EP. In the result, starry flounder appeared to have the better feed preference to the EP, but have the higher feed efficiency and growth performance to the MP.

폴리오바이러스의 분자생물학 (Molecular Miology of the Poliovirus)

  • 최원상
    • 생명과학회지
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    • 제7권4호
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    • pp.392-401
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    • 1997
  • 폴리오바이러스는 바이러스들 중에서도 특히 커기가 작은 바이러스로서 피막(coat)을 둘러싸는 막(envelop) 이 없다. 폴리오바이러스는 (+) 가닥의 단일 RNA 게놈을 갖는데 이는 한 개의 해독판 (open reading frame)을 이용하여 다단백전구체를 만든 후 바이러스 자체의 단백질분해효소에 의해 스스로 잘라져서 궁극적으로느 특이한 기능을 갖는 여러개의 단백질이 된다. P1 다단백질전구체로부터 만들어지는 단백질들은 바이러스의 피막을 구성하는 성분이다. 단백질분해효소인 2A에 의한 최초의 절단은 구조단백질 P1 전구체와 구조단백질이 아닌 P2-P3간을 분리시켜준다. 단백질분해효소 2A는 진핵세포 판독개시인자(translation initiation factor) 4F의 한 subunit인 숙주단백질 p220의 절단에 간접으로 참여한다. 이 단백질의 절단은 캡(cap)에 의존하는 숙주세포의 대부분의 판독을 차단하게 되며 이는 판독에 사용되는 숙주세포의 모든 기구들을 캡에 의존하지 않는 폴리오바이러스 NA 특유의 판독을 위해 전적으로 사용할 수 있게 해준다. 2B, 2C, 2BC 단백질의 기능에 대해서는 많이 알려져 있지 않다. 2B, 2C, 2BC와 3CD 단백질들은 바이러스로 인해 만들어지는 소낭(vesicle)의 복제복합체에 함유되어 있으므로 바이러스의 RNA 복제시 중요한 역할을 함을 암시해준다. 새로이 만들어진 모든 바이러스 RNA는 VPg와 공유결합으로 연결되어 있다. VPg는 3AB로부터 만들어진 아미노산 22개 짜리의 폴리펩타이드이다. 3C와 3CD는 단백질분해소로 다단백질 전구체의 대부분의 절단부위를 잘라준다. 3C단백질은 숙주의 전사인자를 불활성화 시킴으로써 RNA polymer II와 III에 의한 전사를 저해한다. 3D는 RNA의존선RNA 중합효소이다. 폴리오바이러스는 (+)가닥 RNA 바이러스의 일반적인 복제양식을 따른다. 즉 (+) 가닥 RNA는 이와 상보적인 (-)가닥 RNA로 전사되고 이는 다시 (+)가닥 RNA의 합성을 위한 주형으로 사용된다. 폴리오바이러스의 RNA 합성은 세포내막에서 일어나는 데 RNA 복제에 요구되는 주형 RNA와 이때 필요한 단백질들이 어떤 방법으로 세포내막에서 모일 수 있는지는 아직 밝혀진 것이 적다. 바이러스입자의 형성은 세포막의 RNA 복제가 들어가는 데 피막단백질이 (+)가닥 RNA을 인식하는 표지 즉 packaging singal에 대해서는 거의 알려져 있지 않다. 폴리오바이러스 감염 후 첫 바이러스입자가 만들어지기 까는 약 6시간이 소요된다.

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Experimental Infection of Different Tomato Genotypes with Tomato mosaic virus Led to a Low Viral Population Heterogeneity in the Capsid Protein Encoding Region

  • Sihelska, Nina;Vozarova, Zuzana;Predajna, Lukas;Soltys, Katarina;Hudcovicova, Martina;Mihalik, Daniel;Kraic, Jan;Mrkvova, Michaela;Kudela, Otakar;Glasa, Miroslav
    • The Plant Pathology Journal
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    • 제33권5호
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    • pp.508-513
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    • 2017
  • The complete genome sequence of a Slovak SL-1 isolate of Tomato mosaic virus (ToMV) was determined from the next generation sequencing (NGS) data, further confirming a limited sequence divergence in this tobamovirus species. Tomato genotypes Monalbo, Mobaci and Moperou, respectively carrying the susceptible tm-2 allele or the Tm-1 and Tm-2 resistant alleles, were tested for their susceptibility to ToMV SL-1. Although the three tomato genotypes accumulated ToMV SL-1 to similar amounts as judged by semiquantitative DAS-ELISA, they showed variations in the rate of infection and symptomatology. Possible differences in the intra-isolate variability and polymorphism between viral populations propagating in these tomato genotypes were evaluated by analysis of the capsid protein (CP) encoding region. Irrespective of genotype infected, the intra-isolate haplotype structure showed the presence of the same highly dominant CP sequence and the low level of population diversity (0.08-0.19%). Our results suggest that ToMV CP encoding sequence is relatively stable in the viral population during its replication in vivo and provides further demonstration that RNA viruses may show high sequence stability, probably as a result of purifying selection.

Cytotoxic T Lymphocytes Elicited by Dendritic Cell-Targeted Delivery of Human Papillomavirus Type-16 E6/E7 Fusion Gene Exert Lethal Effects on CaSki Cells

  • Wu, Xiang-Mei;Liu, Xing;Jiao, Qing-Fang;Fu, Shao-Yue;Bu, You-Quan;Song, Fang-Zhou;Yi, Fa-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권6호
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    • pp.2447-2451
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    • 2014
  • Human papillomavirus (HPV) is the primary etiologic agent of cervical cancer. Consideration of safety and non human leukocyte antigen restriction, protein vaccine has become the most likely form of HPV therapeutic vaccine, although none have so far been reported as effective. Since tumor cells consistently express the two proteins E6 and E7, most therapeutic vaccines target one or both of them. In this study, we fabricated DC vaccines by transducing replication-defective recombinant adenoviruses expressing E6/E7 fusion gene of HPV-16, to investigate the lethal effects of specific cytotoxic T lymphocytes (CTL) against CaSki cells in vitro. Mouse immature dendritic cells (DC) were generated from bone marrow, and transfected with pAd-E6/E7 to prepare a DC vaccine and to induce specific CTL. The surface expression of CD40, CD68, MHC II and CD11c was assessed by flow cytometry (FCM), and the lethal effects of CTL against CaSki cells were determined by DAPI, FCM and CCK-8 methods. Immature mouse DC was successfully transfected by pAd-E6/E7 in vitro, and the transfecting efficiency was 40%-50%. A DC vaccine was successfully prepared and was used to induce specific CTL. Experimental results showed that the percentage of apoptosis and killing rate of CaSki cells were significantly increased by coculturing with the specific CTL (p <0.05). These results illustrated that a DC vaccine modified by HPV-16 E6/E7 gene can induce apoptosis of CaSki cells by inducing CTL, which may be used as a new strategy for biological treatment of cervical cancer.

Use of Tannery Wastes in the Diet of Broiler

  • Alam, M.J.;Amin, M.R.;Samad, M.A.;Islam, M.A.;Wadud, M.A.
    • Asian-Australasian Journal of Animal Sciences
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    • 제15권12호
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    • pp.1773-1775
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    • 2002
  • Tannery waste contained 90.93% DM, 77.02% CP, 0.77% CF, 2.83% EE, 7.19% ash and 3,450 kcal ME/kg DM. A total of 144 day-old broiler chicks were divided into three dietary groups; $D_1$ (Containing 10% protein concentrate-PC), $D_2$ (Containing 5% PC+5% tannery waste-TW) and $D_3$ (Containing 10% TW) having 3 replicates of 16 chicks in each. The birds were fed broiler starter diet containing 22% CP, 3,000 kcal ME/kg and broiler finisher diet containing 21% CP, 3,100 kcal ME/kg up to 42 days of age, and meat yield traits were measured from the representative birds from each replication to asses the feasibility of using tannery waste in the diet of broiler. Feed intake, live weight, feed conversion efficiency and livability did not differ between diets (p>0.05) but the cost of production and profitability differed significantly (p<0.001). Profitability of D1, D2, and D3 diets were 2.98, 9.90 and 14.04 Taka/kg respectively. Diets did not affect on meat yield traits (p>0.05), except gizzard, shank and feather weight (p<0.01). Gizzard and shank weigh were improved with increasing level of tannery wastes in the diet, hence tannery waste can be used without any harmful effect in the broiler diet.

Effect of combinatorial bone morphogenetic protein 2 and bone morphogenetic protein 7 gene delivery on osteoblastic differentiation

  • Bae, Young;Kim, Kyoung-Hwa;Kim, Su-Hwan;Lee, Chul-Woo;Koo, Ki-Tae;Kim, Tae-Il;Seol, Yang-Jo;Ku, Young;Rhyu, In-Chul;Chung, Chong-Pyoung;Lee, Yong-Moo
    • Journal of Periodontal and Implant Science
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    • 제39권sup2호
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    • pp.279-286
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    • 2009
  • Purpose: Gene therapy (ex vivo) has recently been used as a means of delivering bone morphogenetic proteins (BMPs) to sites of tissue regeneration. In the present study, we investigated the effect of co-transduction of adenoviruses expressing BMP-2 and BMP-7 on osteogenesisof C2C12 cells in vitro. Methods: A replication-defective human adenovirus 5 (Ad5) containing a cDNA for BMPs in the E1 region of the virus (Ad5BMP-2 and Ad5BMP-7) was constructed by in vivo homologous recombination. Functional activity of Ad5BMP-2 and Ad5BMP-7 were evaluated in mouse stromal cells (W20-17cells). C2C12 cells are transduced with various MOI (multiplicity of infection) of Ad5BMP-2 and Ad5BMP-7 to assess most effective and stable titer. Based on this result, C2C12 cells were transduced with Ad5BMP-2 and Ad5BMP-7 alone or by combination. BMPs expression, alkaline phosphatase (ALPase) activity, cell proliferation, and mineralization were assessed. Results: Ad5BMP-2 and Ad5BMP-7 are successfully transduced to W20-17 cells, and secreted BMPs stimulated cell differentiation. Also, C2C12 cells transduced with Ad5BMPs showed expression of BMPs and increased ALPaseactivity. In all groups, cell proliferation was observed over times. At 7days, cells co-transduced with Ad5BMP-2 and Ad5BMP-7 showed lower proliferation than the others. C2C12 cells co-transduced with Ad5BMP-2 and Ad5BMP-7 had greater ALPaseactivity than that would be predicted if effect of individual Ad5BMPs were additive. Little mineralized nodule formation was detected in cells transduced with individual Ad5BMPs. In contrast, Ad5BMP-2 and Ad5BMP-7 combination stimulated mineralization after culturing for 10 days in mineralizing medium. Conclusions: Present study demonstrated that adenoviruses expressing BMPs gene successfully produced BMPs protein and these BMPs stimulated cells to be differentiated into osteoblastic cells. In addition, the osteogenic activity of Ad5BMPs can be synergistically increased by co-transduction of cells with Ad5BMP-2 and Ad5BMP-7.

여중생의 월경 불편감과 식습관 관련 특성 (Menstrual Discomfort and Dietary Habits in Adolescents)

  • 홍경자;김혜원;안혜영
    • Child Health Nursing Research
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    • 제11권3호
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    • pp.330-339
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    • 2005
  • Purpose: This study was done to identify menstrual discomfort and dietary habits, and factors related to the menstrual discomforts. Method: Participants were 320 female middle school students in G city. The Menstrual Discomfort Questionaire(MDQ) and dietary habit lists were used as tools. Results: Factors related to MDQ were found to be the VAS scores (r=.361, p=.002), a mount of menstrual bleeding (r=.131, p=.019), height (r=.134, p=.016), adequacy of meal time (t=7.19, p=.008), consumption of milk & milk products (F=3.20, p=.042) and, hot, salty & irritant foods (f=8.01, p=.000), eating more than 3 kinds of side-dishes with each meal (F=8.32, p=.000), and various protein foods (F=5.15, p=.006). In stepwise regression, 4 variables (VAS scores, height, hot, salty & irritant foods, more than 3 kinds of side-dishes with each meal) explained $20.3\%$ of the variance in the total MDQ scores. Conclusion: Reduction of hot, salty & irritant foods and having more than 3 side-dishes with each meal would be effective in menstrual discomfort management. As well, good and, balanced dietary habits should be encouraged for early adolescent girls. To keep up with the ideal dietary habits, nutritional education & counseling should be continued. In a further study, a replication study with larger samples and more specified nutritional assessment are recommended.

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Expression and Characterization of RNA-dependent RNA Polymerase of Dendrolimus punctatus Tetravirus

  • Zhou, Liang;Zhang, Jiamin;Wang, Xiaochun;Jiang, Hong;Yi, Fuming;Hu, Yuanyang
    • BMB Reports
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    • 제39권5호
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    • pp.571-577
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    • 2006
  • Dendrolimus punctatus tetravirus (DpTV) has been identified as a new member of the genus Omegatetravirus of the family Tetraviridae that may be related serologically to Nudaurelia capensis virus ($N{\omega}V$). To establish the function of DpTV RNA genome and to better understand the mechanism of viral replication, the putative RNA-dependent RNA polymerase (RdRp) domain has been cloned and expressed in Escherichia coli. The recombinant protein was purified on a Ni-chelating HisTrap affinity column and demonstrated to initiate viral RNA synthesis in a primer-independent manner but not by terminal nucleotidyle transferase activity in the presence of $Mg^{2+}$ and RNA template. Mutation of the GDD to GAA interferes with the residues at the polymerase active site and metal ions, and thus renders the polymerase inactive.