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Biochemical Quantitation of PM2 Phage DNA as a Substrate for Endonuclease Assay

  • Joo, Yoo-Jin;Kim, Hee-Ju;Lee, Jae-Yung;Kim, Joon
    • Journal of Microbiology
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    • v.42 no.2
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    • pp.99-102
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    • 2004
  • Bacteriophage PM2 has a closed circular form of double stranded DNA as a genome. This DNA from the phage is a useful source for nick-circle endonuclease assay in the fmol range. Due to difficulties in the maintenance of viral infectivity, storage conditions of the phage should be considered for the puri-fication of PM2 DNA. The proper condition for a short-term storage of less than 2 months is to keep the PM2 phage at 4$^{\circ}C$; whereas the proper condition for a long-term storage of the PM2 phage for over 2 months is to keep it under liquid nitrogen in 7.5 % glycerol. The optimal conditions for a high yield of phage progeny were also considered with the goal to achieve a successful PM2 DNA preparation. A MOI(Multiplicity Of Infection) of 0.03, in which the OD$\sub$600/ of the host bacteria was between 0.3 and 0.5, turned out to be optimal for the mass production of PM2 phage with a burst size of about 214. Considerations of PM2 genome size, and the concentrations and radiospecific activities of purified PM2 DNA, are required to measure the endonuclease activity in the fmol range. This study reports the proper quantitation of radioactivity and the yield of purified DNA based on these conditions.

Germ Cell Apoptosis in the Testis of Transgenic Pigs

  • Chung, Hak-Jae;Kim, Bong-Ki;Ko, Yeoung-Gyu;Woo, Jei-Hyun;Kim, Jeom-Soon;Jung, Jin-Kwan;Chang, Won-Kyong
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.233-233
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    • 2004
  • PURPOSE: Gene expression and apoptosis in testicular germ cells has been demonstrated in many transgenic animals. However, little is known about the transgenic pig and rates of apoptosis during spermatogenesis. METHODS : Morphological and biochemical features of apoptosis reported in other species were used to confirm that the TdT-mediated dUTP Nick end labeling (TUNEL) assay is an acceptable mothos for idendtification and quantification of apoptotic transgenic germ cells in histological tissue section from transgenic pig testis. (omitted)

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Apoptosis of Parthenogenetic Preimplantation Porcine Embryos Activated and Cultured in Different Condition

  • Hwang, In-Sun;Im, Gi-Sun;Kim, Dong-Hoon;Yang, Byoung-Chul;Park, Hyo-Suk;Kim, Se-Woong;Seo, Jin-Sung;Yang, Bo-Suk;Chang, Won-Kyong
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.237-237
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    • 2004
  • Parthenogenesis and culture condition are essential to intracytoplasmic sperm injection and cloning by nuclear transfer. This study investigated apoptosis and in vitro development of parthenogenetic preimplantation porcine embryos. 42∼44 h in vitro matured oocytes derived from a local abattoir were used. Apoptotic cell death was analyzed by using a terminal deoxynucleatidyl transferase mediated deoxyuridine 5-triphoshate nick-end labling (TUNEL) assay. (omitted)

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Styrylpyrone Derivative Induces Apoptosis through the Up-Regulation of Bax in the Human Breast Cancer Cell Line MCF-7

  • Chien, Alvin Lee Teck;Pihie, Azimahtol Hawariah Lope
    • BMB Reports
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    • v.36 no.3
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    • pp.269-274
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    • 2003
  • In the fight against cancer, novel chemotherapeutic agents are constantly being sought to complement existing drugs. Various studies have presented evidence that the apoptosis that is induced by these anticancer agents is implicated in tumor regression, and Bcl-2 family genes play a part in apoptosis following treatment with various stimuli. Here, we present data that a styrylpyrone derivative (SPD) that is extracted from the plant Goniothalamus sp. showed cytotoxic effects on the human breast cancer cell line MCF-7. SPD significantly increased apoptosis in MCF-7 cells, as visualized by phase contrast microscopy and evaluated by the Tdt-mediated dUTP nick end-labeling assay and nuclear morphology. Western blotting and immunostaining revealed up-regulation of the proapoptotic Bax protein expression. SPD, however, did not affect the expression of the anti-apoptotic protein, Bcl-2. These results, therefore, suggest SPD as a potent cytotoxic agent on MCF-7 cells by inducing apoptosis through the modulation of Bax levels.

Neuroprotective Effect of Ginseng radix on ICH-induced Rats

  • Jang, Kwan-Ho;Song, Yun-Kyung;Lim, Hyung-Ho
    • The Journal of Korean Medicine
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    • v.26 no.4
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    • pp.87-97
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    • 2005
  • Backgrounds: Intracerebral hemon-hage is one of the most devastating types of stroke. Ginseng radix, the root of Panax Ginseng, C. A. MEYER (Araliaceae), is one of the most famous medicinal herbs with various therapeutic applications. Objectives: In the present study, the effect of aqueous extract of Ginseng radix on intracerebral hemorrhage-induced neuronal cell death in rats was investigated. Materials and Methods: Step-down avoidance task, Nissl staining, immunohistochemistry for caspase-3 and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay were used for this study. Results: The present results show that hemorrhage-induced lesion volume and apoptotic neuronal cell death in the striatum were significantly suppressed by treatment with Ginseng radix, resulting in enhancement of short-ten-n memory. Conclusions: We have shown that Ginseng radix has a neuroprotective effect on stroke, and aids the recovery from central nervous system sequelae following stroke.

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Acupuncture Suppresses Intrastriatal Hemorrhage-Induced Neuronal Cell Death and Proliferation in Rats

  • Cho, Nam-Hun;Park, Dong-suk;Lee, Jae-Dong
    • The Journal of Korean Medicine
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    • v.24 no.4
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    • pp.127-135
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    • 2003
  • Objectives : Intracerebral hemorrhage (ICH) is one of the most devastating types of stroke. The effect of acupuncture on the intrastriatal hemorrhage-induced neuronal cell death and cell proliferation in rats is examined. Methods : Cell death and cell proliferation in rats was investigated via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay and immunohistochemistry for caspase-3 and 5-bromo-2'-deoxyuridine (BrdU). Results : Results showed that apoptotic cell death in the striatum and cell proliferation in the hippocampal dentate gyrus significantly increased following intrastriatal hemorrhage in rats, and that acupunctural treatment at the Zusanli acupoint suppressed the hemorrhage-induced increase in apoptosis in the striatum and cell proliferation in the dentate gyrus. Conclusions : It is suggested that acupunctural treatment, especially at the Zusanli acupoint, may aid recovery following central nervous system sequelae following ICH.

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Blends of semi-rigid substituted poly(p-phenylene) with BPA-polycarbonate

  • Dijkstra Dirk J.;Karbach Alexander;Malkovich Nick
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.364-364
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    • 2006
  • The rheological properties of Parmax 1200, a new semi-flexible substituted polyphenylene, were investigated. The reported high stiffness of the material was confirmed. The rheological measurements proved that, despite the very high stiffness of the molecules, Parmax showed shear thinning and that, although the viscosity is very high and the melt is highly elastic, the polymer can be extruded in the melt. A worm-like morphology was detected in AFM and TEM. This morphology could explain the reported mechanical and rheological behaviour. The compatibility with flexible chain polymers (e.g. polycarbonate) could also be explained by the worm-like morphology.

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A Small Cryptic Plasmid pZMO1 of Zymomonas mobilis ATCC10988

  • Kang, Hyung-Lyun;Kang, Hyen-Sam
    • Genomics & Informatics
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    • v.1 no.1
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    • pp.55-60
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    • 2003
  • The nucleotide sequence of pZMO1, a small cryptic plasmid of Zymomonas mobilis ATCC10988 was determined. Analysis of 1,680 bp of sequence revealed $69\%$ identity with Shigella sonnei plasmid, pKYM and $61\%$ identity with Nostoc sp. ss DNA replicating plasmid. Analysis of a deduced amino acid sequence of an orf of pZMO1 revealed $75\%$ identity and $90\%$ similarity with the repA gene of Synechocystis sp. plasmid pCA2.4. The upstream region of the repA gene of pZMO1 possesses six directed repeat sequences and two inverted repeat sequences at downstream of the IR consensus sequence of nick region of rolling circle replication (RCR) plasmid. A typical terminator hairpin structure was found at the downstream region of repA gene. Degradation of single-stranded plasmid DNA by S1 nuclease was detected by Southern hybridization. It suggests that pZMO1 replicates by a rolling circle mechanism in Z. mobilis ATCC10988 cells.

Biological Activity of Extracts from Zea mays L. and Pinus densiflora L. (옥수수(Zea mays L.)와 소나무(Pinus densiflora L.) 추출물의 생물학적 활성)

  • ;Soul Chun;Nick E. Christians
    • Asian Journal of Turfgrass Science
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    • v.12 no.4
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    • pp.203-210
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    • 1998
  • Environmental concerns arising from synthetic herbicides in plant management systems have led to an interest in plant-derived compounds as natural herbicides. Inhibitory effects of compounds extracted with 50% methanol from corn (Zea mays L.) and pine (Pinus densiflora L.) were evaluated on large crabgrass (Digitaria sanguinalis (L.) Scop.), annual bluegrass Poa annua L.), radish (Raphanus sativus L.), and perennial ryegrass (Lolium perenne L.) The aqueous extracts inhibited seed germination and had postemergence activity on the four species. The stability of biological activity of corn grain, stover, and root extracts was not affected by heating to $135^{\circ}C$ or freezing/thawing treatments when applied at levels above 0.25kg m(sup)-2 based on dry weights of powders before extraction. Heating reduced the activity of pine litter and bark extracts at all levels except the highest application level but had little effect on pine needle extracts.

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A Development of the Noise Quality Checking System EOL of the 6th Speed Automatic Transmission in the USA (북미 6속 자동변속기 공장 EOL NVH 시스템 개발에 대한 연구)

  • Lee, Hyun-Ku;Kim, Moo-Suk;Hwang, Seon-Yang;Kang, Koo-Tae;Lee, Tae-Hwi
    • Proceedings of the Korean Society for Noise and Vibration Engineering Conference
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    • 2011.04a
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    • pp.731-732
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    • 2011
  • This paper shows a whole noise quality checking system for the new developed 6th speed automatic transmission manufactured in USA. Due to demand for manufacturing of the high NVH quality transmissions in the mass product line, some special checking system called EOL is adapted. This EOL system is using vibration analysis technologies. Through adapting vibration analysis technologies and functions likewise RMS, Crest Factor, Harmonics, Peak, Band, and Order tracking, various noise problems caused by wear, nick, and deformations could be successfully detected and predicted. Therefore, automatic transmissions manufactured in the USA could get high NVH quality.

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