To understand the molecular structure of Korean garlic viruses, cDNA cloning of virus genomic RNA was attempted. Virus particles were isolated from virus-infected garlic leaves and a cDNA library was constructed from garlic virus RNA. One of these clones, S81, selected by random sequencing has been identified as a member of potexvirus group other than potyvirus and carlavirus. The clone is 873 bp long contains most of the coat protein (CP) coding region and 3'-noncoding region including poly(A) tail. A putative polyadenylation signal sequence (AAUAAA) and the hexanucleotide motif (ACUUAA), a replicational cis-acting element conserved in the 3'-noncoding region of potexvirus RNAs are noticed. The clone S81 shows about 30-40% identity in both nucleotide and amino acid sequences with CPs of potexviruses. The genome size of the virus was analysed to be 7.46 knt by Northern blot analysis, which was longer than those of other potexviruses. The open reading frame encoding CP was expressed as a fusion protein (S81CP) in Escherichia coli and the recombinant protein was purified by immobilized metal binding affinity chromatography. Polyclonal antibody was raised against S81CP in rabbit to examine the occurrence of garlic potexvirus in Korean garlic plants by immunoblot analysis. Two virus protein bands of Mr 27,000 and 29,000 from garlic leaf extract of various cultivars reacted with the antibody. It was shown that Mr 27,000 band might not be a degradation product of Mr 29,000 band, suggesting that two types of potexvirus different in size of coat protein could exist in Korean garlic plants.
RNase E is an essential Escherichia coli endoribonuclease that plays a major role in the decay and processing of a large fraction of RNAs in the cell and expression of N-terminal domain consisted of 1-498 amino acids (N-Rne) is sufficient to support normal cellular growth. By utilizing these properties of RNase E, we developed a genetic system to screen for amino acid substitutions in the catalytic domain of the protein (N-Rne) that lead to various phenotypes. Using this system, we identified three kinds of mutants. A mutant N-Rne containing amino acid substitution in the S1 domain (I6T) of the protein was not able to support survival of E. coli cells, and another mutant N-Rne with amino acid substitution at the position 488 (R488C) in the small domain enabled N-Rne to have an elevated ribonucleolytic activity, while amino acid substitution in the DNase I domain (N305D) only enabled N-Rne to support survival of E. roli cells when the mutant N-Rne was over-expressed. Analysis of copy number of ColEl-type plasmid revealed that effects of amino acid substitution on the ability of N-Rne to support cellular growth stemmed from their differential effects on the ribonucleolytic activity of N-Rne in the cell. These results imply that the genetic system developed in this study can be used to isolate mutant RNase E with various phenotypes, which would help to unveil a functional role of each subdomain of the protein in the regulation of RNA stability in E. coli.
Yoo, Seon A;Kim, Ok Kyung;Nam, Da-Eun;Kim, Yongjae;Baek, Humyoung;Jun, Woojin;Lee, Jeongmin
Journal of the Korean Society of Food Science and Nutrition
/
v.43
no.2
/
pp.216-223
/
2014
Curcuma longa L. (CL) is a well known traditional medicinal plant that is also used in curries and mustards as a coloring and flavoring agent. However, CL is not usually used as a food source due to its bitter taste. We investigated the immunomodulatory effect of CL fermented by Aspergillus oryzae (FCL) on RAW 264.7 cells. FCL was extracted with cold water (CW), hot water (HW), 20% ethanol (20% EtOH) and 80% ethanol (80% EtOH), after which its effects on phagocytic activity, tumor necrosis factor-alpha (TNF-${\alpha}$), nitric oxide (NO) production, natural killer (NK) cell activity and mRNA expression of LP-BM5 eco were investigated. Phagocytic activity was increased in HW and 20% EtOH when compared to the control. The secretion of nitric oxide (NO) from RAW 264.7 cells did not change significantly relative to the control. However, TNF-${\alpha}$ was significantly increased by the addition of FCL extracts. Moreover, FCL 20% ethanol extract showed a four fold increase in NK cell cytotoxity relative to the control group. Finally, we observed suppressed mRNA expression of LP-BM5 eco in FCL extracts, especially in the 20% ethanol extracts group. These results indicate that the FCL extracts can be used as a functional material due to their effective immunomodulating activities.
The possibility of inadvertent introduction of therapeutic gene expressing viral vectors has raised safety concerns about germ-line infection. Particularly, for indications such as prostate cancer and ovarian cancer, the proximity of the point of viral administration to organs of the reproductive system raises concerns regarding inadvertent germ-line transmission of genes carried by the virus vector. To evaluate the safety of in vivo adenovirus mediated gene transfer, we explored the biodistribution, persistance and potential germ-line transmission of p53-expressing adenovirus (Ad-CMV-p53). Both male and female Balb/c mice were injected with $1{\times}10^9$ PFU of Ad-CMV-p53. The PCR analysis showed that there were detectable vector sequences in liver, kidney, spleen, seminal vesicle, epididymis, prostate, ovary, and uterus. The RT-PCR analysis for detecting inserted gene, p53 showed that Ad-CMV-p53 viral RNA were present in spleen, prostate and ovary. Direct injected male and female mice of adenovirus vector into testis and ovary were mated and their of offspring were evaluated for germ-line transmission of the adenoviral vector. The PCR and RT-PCR analysis showed no evidence of germline transmission, although vector sequences were detected in DNA extracted from gonadal tissues. Real-time PCR result confirmed a significant decrease of adenovirus in gonad tissues 1 week after injection. We have also analysed the cell specific localization of viral DNA in gonad tissues by using in-situ PCR. Positive signals were detected in interstitial tissue but not in seminiferous tubule in sperm. In the case of ovary, adenovirus signal were localized to the stromal tissue, but no follicular signals were observed. Together, these data provide strong evidence that the risk of the Inadvertent germ-line transmission of vector sequences following intraperitoneal or direct injection into genito-urinary system of adenovirus is extremely low.
The purpose of this study is to evaluate the effects of surface treatment and composition of reinforcement material on fracture strength of fiber reinforced composite inlay bridges. The materials used for this study were I-beam, U-beam TESCERA ATL system and ONE STEP(Bisco, IL, USA). Two kinds of surface treatments were used; the silane and the sandblast. The specimens were divided into 11 groups through the composition of reinforcing materials and the surface treatments. On the dentiform, supposing the missing of Maxillary second pre-molar and indirect composite inlay bridge cavities on adjacent first pre-molar disto-occlusal cavity, first molar mesio-occlusal cavity was prepared with conventional high-speed inlay bur. The reinforcing materials were placed on the proximal box space and build up the composite inlay bridge consequently. After the curing, specimen was set on the testing die with ZPC. Flexural force was applied with universal testing machine (EZ-tester; Shimadzu, Japan). at a cross-head speed of 1 mm/min until initial crack occurred. The data was analyzed using one-way ANOVA/Scheffes post-hoc test at 95% significance level. Groups using I-beam showed the highest fracture strengths (p<0.05) and there were no significant differences between each surface treatment (p>0.05) Most of the specimens in groups that used reinforcing material showed delamination. 1. The use of I-beam represented highest fracture strengths (p<0.05) 2. In groups only using silane as a surface treatment showed highest fracture strength, but there were no significant differences between other surface treatments (p>0.05). 3. The reinforcing materials affect the fracture strength and pattern of composites inlay bridge. 4 The holes at the U-beam did not increase the fracture strength of composites inlay bridge.
Thanks to recent advances in next-generation sequencing (NGS) technology, diverse livestock species have been dissected at the genome-wide sequence level. As for cattle, there are currently four Korean indigenous breeds registered with the Domestic Animal Diversity Information System of the Food and Agricultural Organization of the United Nations: Hanwoo, Chikso, Heugu, and Jeju Heugu. These native genetic resources were recently whole-genome resequenced using various NGS technologies, providing enormous single nucleotide polymorphism information across the genomes. The NGS application further provided biological such that Korean native cattle are genetically distant from some cattle breeds of European origins. In addition, the NGS technology was successfully applied to detect structural variations, particularly copy number variations that were usually difficult to identify at the genome-wide level with reasonable accuracy. Despite the success, those recent studies also showed an inherent limitation in sequencing only a representative individual of each breed. To elucidate the biological implications of the sequenced data, further confirmatory studies should be followed by sequencing or validating the population of each breed. Because NGS sequencing prices have consistently dropped, various population genomic theories can now be applied to the sequencing data obtained from the population of each breed of interest. There are still few such population studies available for the Korean native cattle breeds, but this situation will soon be improved with the recent initiative for NGS sequencing of diverse native livestock resources, including the Korean native cattle breeds.
This study was done to evaluate whether there were any differences in microleakage of class V composite restorations according to restoration site and cavity size. Total sixty-four restorations were made in molar teeth using Esthet-X. Small ($2\;{\times}\;2\;{\times}\;1.5\;mm$) and large ($4{\times}2{\times}1.5\;mm$) restorations were made at the buccal/lingual surface and the proximal surface each. After 1,000 times of thermocycling ($5^{\circ}\;-\;55^{\circ}C$), resin replica was made and the percentage of marginal gap to the whole periphery of the restoration was estimated from SEM evaluation. Thermocycled tooth was dye penetrated with $50\%$ silver nitrate solution. After imbedding in an auto-curing resin, it was serially ground with a thickness of 0.25 mm. Volumetric microleakage was estimated after reconstructing three dimensionally. Two-way ANOVA and independent T-test for dye volume, Mann-Whitney U test for the percentage of marginal gap, Spearman's rho test for the relationship between two techniques were used, The results were as follows : 1. The site and size of the restoration affected on the microleakage of restoration. Namely, much more leakage was seen in the proximal and the large restorations rather than the buccal/lingual and the small restorations. 2. Close relationship was found between two techniques (Correlation coefficient = 0.614/ P = 0.000). Within the limits of this study, it was noted that proximal and the large restorations leaked more than buccal/lingual and the small restorations. Therefore, it should be strictly recommended large exposure of margins should be avoided by reducing unnecessary tooth reduction.
This study was conducted to examine the effects of electric stimulation conditions on in vitro developmental ability of procine embryos after somatic cell nuclear transfer, The porcine ear cell was cultured in vifro for confluency in serum-starvation condition (TCM-199+0.5% FBS) for cell confluency. The zona pellucida of IVM oocytes were partially drilled using laser system. Single somatic cell was individually transferred into the enucleated oocyte. The reconstructed embryos were electrically fused with 0.3M mannitol. After electric fusion, the embryos were activated and cultured in NCSU-23 medium containing 10% FBS at 39$^{\circ}C$, 5% $CO_2$ in air for 6 to 8 days. Nuclear transferred(NT) oocytes which fused at a field strength of 1.90kv/cm showed a higher (P<0.05) fusion rate(49.5%, 50/101) compared to 2.10 kv/cm(25.8%, 24/93) or 2.50kv/cm(30.3%, 27/89). After electric activation, the cleavage rate of NT embryos was 48.0(24/50), 66.6(16/24) and 70.3% (19/27), respectively and these were not different. There was no significant difference in fusion rate by duration and pulse of electric stimulation. In cleavage rate, however, more NT embryos(76.3%, 45/59) cleaved at 60 $\mu$sec twice than other embryos(49.1 to 56.5%) with different conditions of electric stimulation(P<0.05). NT embryos activated at a field strength of 1.50kv/cm showed a higher developmental rate(9.8%, 5/51) than those embryos activated at 1.25kv/cm(0%) or parthenotes(6.4%, 7/109). These results suggest that some factors such as field strength, duration and pulse of electric stimulation could be affected to in vitro developmental ability of nuclear transplanted porcine embryos.
Journal of the korean academy of Pediatric Dentistry
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v.50
no.1
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pp.113-120
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2023
Restoring composite resins with the optimal shades for all primary teeth is a great challenge for pediatric dentists. A newly developed single-shade composite resin can exhibit a color similar to that of the surrounding tooth structure based on the structural color phenomenon. This study aims to evaluate the color adjustment potential (CAP) of a single-shade composite resin compared to conventional multi-shade composite resins in primary teeth. A single-shade composite resin and two conventional multi-shade composite resins were included in this study. Two types of specimens, a single specimen and a dual specimen, were evaluated. For single specimens, duplications of the primary second molar denture teeth were made using experimental composite resins. For dual specimens, cavities were prepared on the buccal surfaces of extracted primary second molars and restored with experimental composite resins. The L*, a*, and b* values were measured using a colorimeter for the extracted teeth and specimens. The mean ΔEab* values for single and dual specimens and CAP were calculated. Bonferroni post-hoc analysis was performed to confirm the statistical significance between the ΔEab* and CAP values of the experimental resins. Among the single specimens, the single-shade composite resin showed significantly higher ΔESingle compared to other composite resins (p < 0.0167). There was no significant difference between ΔEDual for all experimental resins. The single-shade composite resin showed highest CAP compared to other multi-shade composite resins. A single-shade composite resin exhibited the most prominent color adaptability compared to other conventional multi-shade composite resins for primary second molars. A single-shade composite resin can simplify shade matching and provide esthetic outcomes for the restoration of primary second molars.
This paper take a media archaeological approach to cinema transformed into a narrative medium during its transitional period, 1903-1915. To accomplish this, I will explore the question of as which narrative medium cinema was imagined and also how it was institutionalized as a narrative medium with authorship. I will explain that the imaginary and real ideas and changes on cinema resonated with each other on the foundation of its technological aspects such as indexicality, 23 frames/sec. and montage. It was during the transitional period that cinema was transformed from a medium representing spectacle to a medium of narration. The establishment of the American film copyright law in 1912 was an institutional, real outcome from the contemporary understanding of cinema as a narrative medium. At the same time, various ideas emerged that led to imagining of cinema as a complete narrative medium, incomparable to any other. From a media archaeological perspective, the imaginary ideas of media resonate with their actual course of development. These imaginary ideas are not just imaginary, but rather reflect the contemporary desire for the medium. This paper looks into the transitional period based on this media archaeological point of view. To this end, this paper will briefly introduce the notion of media archaeology as a media theory and then discuss Eric Kluitenberg's concept of 'an archaeology of imaginary media' and its methodologies. Second, it will explore literary and cinematic imagining of cinema as a powerful medium of storytelling, while discussing the ways in which cinema's technological characteristics played a decisive role in these imaginings. Also to show the techno-deterministic role of cinema in the real world, this paper will explore how its technological characteristics were considered as an important element in the processes through which America's first motion picture copyright was institutionalized in 1912 after two historical copyright cases: one is Edison v. Lubin in 1903 and Kalem v. Harper Brothers in 1909. Ultimately, this paper will lead us to an understanding of the history of cinema as a medium and its developments in more multi-layed way, as communication between the real and imaginary, and give us perspectives toward what cinema is.
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