• Title/Summary/Keyword: in vitro techniques

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Comparison of implant stability measurements between a resonance frequency analysis device and a modified damping capacity analysis device: an in vitro study

  • Lee, Jungwon;Pyo, Se-Wook;Cho, Hyun-Jae;An, Jung-Sub;Lee, Jae-Hyun;Koo, Ki-Tae;Lee, Yong-Moo
    • Journal of Periodontal and Implant Science
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    • v.50 no.1
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    • pp.56-66
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    • 2020
  • Purpose: A stability-measuring device that utilizes damping capacity analysis (DCA) has recently been introduced in the field of dental implantology. This study aimed to evaluate the sensitivity and reliability of this device by measuring the implant stability of ex vivo samples in comparison with a resonance frequency analysis (RFA) device. Methods: Six implant beds were prepared in porcine ribs using 3 different drilling protocols to simulate various implant stability conditions. Thirty-six pork ribs and 216 bone-level implants measuring 10 mm in height were used. The implant beds were prepared using 1 of the following 3 drilling protocols: 10-mm drilling depth with a 3.5-mm-diameter twist drill, 5-mm drilling depth with a 4.0-mm-diameter twist drill, and 10-mm drilling depth with a 4.0-mm-diameter twist drill. The first 108 implants were external-connection implants 4.0 mm in diameter, while the other 108 implants were internal-connection implants 4.3 mm in diameter. The peak insertion torque (PIT) during implant placement, the stability values obtained with DCA and RFA devices after implant placement, and the peak removal torque (PRT) during implant removal were measured. Results: The intraclass correlation coefficients (ICCs) of the implant stability quotient (ISQ) results obtained using the RFA device at the medial, distal, ventral, and dorsal points were 0.997, 0.994, 0.994, and 0.998, respectively. The ICCs of the implant stability test (IST) results obtained using the DCA device at the corresponding locations were 0.972, 0.975, 0.974, and 0.976, respectively. Logarithmic relationships between PIT and IST, PIT and ISQ, PRT and IST, and PRT and ISQ were observed. The mean absolute difference between the ISQ and IST values on a Bland-Altman plot was -6.76 (-25.05 to 11.53, P<0.05). Conclusions: Within the limits of ex vivo studies, measurements made using the RFA and DCA devices were found to be correlated under a variety of stability conditions.

Establishment of efficient Alstromeria callus induction system using node culture and various hormones (마디배양과 다양한 호르몬을 이용한 효율적인 알스트로메리아 캘러스 유도 시스템 체계 확립)

  • Yang, Hwan Rae;Lee, Sang Hee;Kim, Jong Bo
    • The Journal of the Convergence on Culture Technology
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    • v.5 no.1
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    • pp.413-416
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    • 2019
  • Alstroemeria (Alstroemeriaceae) is one of the most important cut flowers in international market. Especially, characteristics like long vase-life, various colors, tolerance to low temperature and a low energy requirement during cultivation have stimulated this success. Because of its characteristics such as low multiplication rates, time-consuming process and high risk of carrying viral disease, in vitro propagation techniques based on rhizome meristems culture have been developing nowadays. The callus induction has various cultivation sites compared with the direct plant generation method, and if the callus is maintained well, the plant differentiation can be performed simultaneously while maintaining the callus, so that it can be used for mass proliferation. In this study, we tested various hormones and cultivars for efficient callus induction. As a result of culturing between the nodes and the internodes, the callus began to be formed after 8 weeks, and the calli incidence in the nodes was higher than that between the internodes. Also, in the comparison of 2,4-D and picloram, the callus incidence rate was up to 2 times higher in the medium treated with 2,4-D. Using these results, it is thought that it will help establish the system of mass propagation system of Alstroemeria and cultivate new varieties.

Glycosyltransformation of ginsenoside Rh2 into two novel ginsenosides using recombinant glycosyltransferase from Lactobacillus rhamnosus and its in vitro applications

  • Wang, Dan-Dan;Kim, Yeon-Ju;Baek, Nam In;Mathiyalagan, Ramya;Wang, Chao;Jin, Yan;Xu, Xing Yue;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.48-57
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    • 2021
  • Background: Ginsenoside Rh2 is well known for many pharmacological activities, such as anticancer, antidiabetes, antiinflammatory, and antiobesity properties. Glycosyltransferases (GTs) are ubiquitous enzymes present in nature and are widely used for the synthesis of oligosaccharides, polysaccharides, glycoconjugates, and novel derivatives. We aimed to synthesize new ginsenosides from Rh2 using the recombinant GT enzyme and investigate its cytotoxicity with diverse cell lines. Methods: We have used a GT gene with 1,224-bp gene sequence cloned from Lactobacillus rhamnosus (LRGT) and then expressed in Escherichia coli BL21 (DE3). The recombinant GT protein was purified and demonstrated to transform Rh2 into two novel ginsenosides, and they were characterized by nuclear magnetic resonance (NMR) techniques and evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assay. Results: Two novel ginsenosides with an additional glucopyranosyl (6→1) and two additional glucopyranosyl (6→1) linked with the C-3 position of the substrate Rh2 were synthesized, respectively. Cell viability assay in the lung cancer (A549) cell line showed that glucosyl ginsenoside Rh2 inhibited cell viability more potently than ginsenoside Rg3 and Rh2 at a concentration of 10 μM. Furthermore, glucosyl ginsenoside Rh2 did not exhibit any cytotoxic effect in murine macrophage cells (RAW264.7), mouse embryo fibroblasts cells (3T3-L1), and skin cells (B16BL6) at a concentration of 10 μM compared with ginsenoside Rh2 and Rg3. Conclusion: This is the first report on the synthesis of two novel ginsenosides, namely, glucosyl ginsenoside Rh2 and diglucosyl ginsenoside Rh2 from Rh2 by using recombinant GT isolated from L. rhamnosus. Moreover, diglucosyl ginsenoside Rh2 might be a new candidate for treatment of inflammation, obesity, and skin whiting, and especially for anticancer.

Breeding of Doritaenopsis 'Hwasu 5205' with Vivid Red and Large Flowers (선명한 적색 대륜계 호접란 '화수 5205' 육성)

  • Lim, Ki-Byung;Kim, Hong-Yul;Park, No-Eun;Son, Beung-Gu;Yun, Suk-Young
    • Horticultural Science & Technology
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    • v.33 no.6
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    • pp.941-946
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    • 2015
  • A new Doritaenopsis cultivar 'Hwasu 5205' was bred by Kyungpook National University, Korea, which produces young plants through tissue culture techniques. The new cultivar 'Hwasu 5205', showing the phenotype of vivid red and large flower type characteristics, was derived from crossing between Phalaenopsis Happy Valentine and Doritaenopsis Happy Rose. An elite individual, number '02-05-205' later named as 'Hwasu 5205', was selected among about 300 individual progenies after more than 2 years of intensive selection covering vegetative and flowering distinctiveness. In year 2004-2005, 1st and 2nd characteristic analyses were carried out through performance and uniformity tests. 'Hwasu 5205' produces vivid red (RHS #PN78B) flowers of i ncurved type with large size, of 9.2 and 12.0 cm in flower height and width, respectively. Leaves of 'Hwasu 5205' grow horizontally and are about 24.3cm in length and 8.5cm in width, respectively. This cultivar possesses no genetic variation. It can be propagated rapidly in vitro and is easy to grow due to its vigorous growth habit. 'Hwasu 5205' was registered (Reg. #: 2915) to Korea Seed & Variety Service (KSVS) on 1st December, 2009 and the PBR(plant breeder's right)is currently controlled by Sangmiwon Orchid Company, Korea.

Real-time Nucleic Acid Sequence Based Amplification (Real-time NASBA) for Detection of Norovirus

  • Lee, In-Soo;Choi, Dong-Hyuk;Lim, Jae-Won;Cho, Yoon-Jung;Jeong, Hye-Sook;Cheon, Doo-Sung;Bang, Hye-Eun;Jin, Hyun-Woo;Choi, Yeon-Im;Park, Sang-Jung;Kim, Sung-hyun;Lee, Hye-Young;Kim, Tae-Ue
    • Biomedical Science Letters
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    • v.17 no.3
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    • pp.191-196
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    • 2011
  • Noroviruses (noroV) are the major cause of nonbacterial gastroenteritis in humans worldwide. Since noroV cannot yet be cultured in vitro and their diagnosis by electron microscopy requires at least $10^6$ viral particles/g of stool a variety of molecular detection techniques represent an important step towards the detection of noroV. In the present study, we have applied real-time nucleic acid sequence-based amplification (real-time NASBA) for simultaneous detection of NoroV genogroup I (GI) and genogroup II (GII) using standard viral RNA. For real-time NASBA assay which can detected noroV GI and GII, a selective region of the genes encoding the capsid protein was used to design primers and genotype-specific molecular beacon probes. The specificity of the real-time NASBA using newly designed primers and probes were confirmed using standard viral RNA of noroV GI and GII. To determine the sensitivity of this assay, serial 10-fold dilutions of standard viral RNA of noroV GI and GII were used for reverse transcription polymerase chain reaction (RT-PCR) and real-time NASBA. The results showed that while agarose gel electrophoresis could detect RT-PCR products with 10 pg of standard viral RNA, the real-time NASBA assay could detect 100 fg of standard viral RNA. These results suggested that the real-time NASBA assay has much higher sensitivity than conventional RT-PCR assay. This assay was expected that might detect the viral RNA in the specimens which could have been false negative by RT-PCR. There were needed to perform real-time NASBA with clinical specimens for evaluating accurate sensitivity and specificity of this assay.

Comparison among the Sperm Preparation Methods on the Human Spermatozoa (인간정자에 있어서 정자처리법의 비교)

  • Pang, Myung-Geol;Chung, Ku-Min;Shin, Chang-Jae;Kim, Jung-Gu;Moon, Shin-Yong;Chang, Yoon-Seok;Lee, Jin-Yong;Lee, Sang-Hoon;Chung, Yung-Chai;Kim, Chang-Keun
    • Clinical and Experimental Reproductive Medicine
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    • v.20 no.2
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    • pp.107-115
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    • 1993
  • Procedures to separate motile. normal & motile and acrosome-reacted sperm with high efficiency have clinical application in Assisted Reproductive Technology in terms of increasing the probability of fertilization by a normal sperm and subsequent normal embryonic development. This study evaluated the effects of 10 sperm preparation techniques [Swim-up from a washed pellet (SU). Swim-up from semen (SO). Continuous Percoll Gradients I (PIC). Discontinuous Percoll Gradients I (PID). Continuous Percoll Gradients II(P II C). Discontinuous Percoll Gradients II(P II D), SpermPrep (SFC). Wang's tube (WT). Albumin Gradients (AG), Low temperature capacitation (LTC)] on motility (%), normal morphology (%), motile sperm recovery rate(%). morphologically normal & motile sperm recovery rate (%), true acrosome reaction (%) and fertilizing ability. A P II D proved to be an effective means of separating morphologically normal & motile sperm. Our results indicated the P II D has advantages as compared with other methods in terms of recovery rate. enhancement of motility and normal morphology. And a LTC seems to be an effective means of enhancing the true acrosome reaction and fertilizing ability. These results suggest that the combined method of LTC and P II D for separation of morphologically normal & motile sperm and acrosome reacted sperm may be a useful procedure for intrauterine insemination and in vitro fertilization in the management of male factor infertility as well as for isolation of subpopulation of sperm for basic research.

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The TNF Receptor Expressions in Cancer Cells Transfected with TNF-$\alpha$ cDNA Using Retroviral Vector (Retroviral vector를 이용한 종양괴사인자 (TNF-$\alpha$) 유전자 이입 암세포에서 종양괴사인자 수용체의 발현)

  • Lee, Hyuk-Pyo;Yoo, Chul-Gyu;Kim, Young-Whan;Shim, Young-Soo;Han, Sung-Koo
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.6
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    • pp.1271-1284
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    • 1997
  • Background : Tumor necrosis factor(TNF) has been considered as an important candidate for cancer gene therapy based on its potent anti-tumor activity. However, since the efficiency of current techniques of gene transfer is not satisfactory, the majority of current protocols is aiming the in vitro gene transfer to cancer cells and re-introducing genetically modified cancer cells to host. In the previous study, it was shown that TNF-sensitive cancer cells transfected with TNF-$\alpha$ cDNA would become highly resistant to TNF, and the probability was shown that the acquired resistance to TNF might be associated with synthesis of some protective protein. Understanding the mechanisms of TNF-resistance in TNF-$\alpha$ cDNA transfected cancer cells would be an important step for improving the efficacy of cancer gene therapy as well as for better understandings of tumor biology. This study was designed to evaluate whether the levels of TNF receptor mRNA expression and soluble TNF receptor release from cancer cells are changed after TNF-$\alpha$ cDNA transfection. Method : We transfected TNF-$\alpha$ c-DNA to WEHI164(murine fibrosarcoma cell line), NCI-H2058(human mesothelioma cell line), A549(human non-small cell lung cancer cell line), ME180(human cervix cancer cell line) cells using retroviral vector(pLT12SN(TNF)) and confirm the expression of TNF with PCR, EUSA, MTT assay. Then we determined the TNF resistance of TNF-$\alpha$ cDNA transfected cells(WEHI164-TNF, NCIH2058-TNF, A549-TNF, ME180-TNF) and evaluated the TNF receptor mRNA expression with Northern blot analysis and soluble TNF receptor release with EUSA. Results : The TNF receptor mRNA expressions of parental cells and genetically modified cells were not significantly different. The soluble TNF receptor levels of media from genetically modified cells were lower than those from parental cells. Conclusion : The acquired resistance to TNF after TNF-$\alpha$ cDNA transfection may not be associated with the change in the TNF receptor and the soluble TNF receptor expression.

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THE EFFECT OF DIFFERENT CURING MODES ON COMPOSITE RESIN/DENTIN BOND STRENGTH IN CLASS ICAVITIES (1급 와동에서 상아질과 복합레진의 결합강도에 대한 중합방법의 효과)

  • Baek, Shin-Young;Cho, Young-Gon;Song, Byeong-Choon
    • Restorative Dentistry and Endodontics
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    • v.33 no.5
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    • pp.428-434
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    • 2008
  • The purpose of this study was to compare the microtensile bond strength in Class I cavities associated with different light curing modes of same light energy density. Occlusal enamel was removed to expose a flat dentin surface and twenty box-shaped Class I cavities were prepared in dentin. Single Bond (3M Dental product) was applied and Z 250 was inserted using bulk technique. The composite was light-cured using one of four techniques, pulse delay (PD group), soft-start (SS group), pulse cure (PC group) and standard continuous cure (CC group). The light-curing unit capable of adjusting time and intensity (VIP, Bisco Dental product) was selected and the light energy density for all curing modes was fixed at $16J/cm^2$. After storage for 24 hours, specimens were sectioned into beams with a rectangular cross-sectional area of approximately $1mm^2$ Microtensile bond strength $({\mu}TBS)$ test was per- formed using a univel·sal testing machine (EZ Test, Shimadzu Co.). The results were analyzed using oneway ANOVA and Tukey's test at significance level 0.05. The ${\mu}TBS$ of PD group and SS group was higher than that of PC group and CC group. Within the limitations of this in vitro study, modification of curing modes such as pulse delay and soft start polymerization can improve resin/dentin bond strength in Class I cavities by controlling polymerization velocity of composite resin.

Breeding of Phalaenopsis 'SM 333' with Mini Multiple Flower Formation (소형 다화 분지성 호접란 'SM 333' 육성)

  • Park, No Eun;Son, Beung Gu;Kim, Hong Yul;Lim, Ki-Byung
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.149-154
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    • 2015
  • A new Phalaenopsis cultivar 'SM 333' was bred by Sangmiwon Orchid, Korea, which produces young plants through tissue culture techniques. The new cultivar 'SM 333', showing the phenotype of multiflora with pink color and small, multibranching-type characteristics, was derived from crossing between Phalaenopsis 'Odoriko' and 'Be Tris'. An elite individual number '02-03-33' later termed 'SM 333' was selected among about 300 individual progenies, based on an intensive selection process covering vegetative and flowering distinctiveness over more than 2 years. In year 2004-2005, the 1st and 2nd characteristic analyses were carried out through performance and uniformity tests. 'SM 333' shows flower color that is bright clean pink (RHS # RP69D) and flower shape that is formal type with 5.0 and 5.8 cm in flower height and width, respectively. 'SM 333' is regarded as raceme flower type suitable for the small casual flower market. The leaves of 'SM 333' grow horizontally and about 20.8 cm in length and 6.5 cm in width. This cultivar also possesses no genetic variation, and is amenable to fast in vitro propagation and easy growth due to its vigorous growth habit. This 'SM333' was registered (Reg. # 2916) with Korea Seed & Variety Service (KSVS) on 1st December, 2009, and the plant breeder's right is currently controlled by Sangmiwon Orchid Company, Korea.

In Vitro Antitumor Activity and Nephrotoxicity of the Novel Platinum(II) Coordination Complex Containing Cis-dach/Diphosphine (새로운 Platinum(II)Complex ([Pt(II)(cis-dach)(DPPP)].$(NO_3)_2$의 항암효과 및 신독성)

  • Jung, Jee-Chang;Yim, Sung-Vin;Park, Seung-Joon;Chung, Joo-Ho;Ko, Kye-Chang;Chang, Sung-Goo;Rho, Young-Soo
    • The Korean Journal of Pharmacology
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    • v.32 no.1
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    • pp.93-102
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    • 1996
  • Platinum coordination complexes are currently one of the most compounds used in the treatment of solid tumors. However, its use is limited by severe side effects such as nephrotoxicity. Our platinum-based drug discovery program is aimed at developing drugs capable of diminishing toxicity and broadening the clinical spectrum of activity of cisplatin. We synthesized new Pt(II) complex analogue containing 1,2-diaminocyclohexane (dach) as carrier ligand and 1,3-bis(diphenyl phosphino)propane (DPPP) as a leaving group. Furthermore, nitrate was added to improve the solubility. A new series of PC-1 [Pt(cis-dach) (DPPP)]. $2NO_3_2$ was synthesized and characterized by their elemental analysis and by various spectroscopic techniques [infrared (IR), $^{13}carbon$ nuclear magnetic resonance (NMR)]. PC-1 was demonstrated acceptable antitumor activity aganist SKOV -3, OVCAR-3 human ovarian adenocarcinomacells and significant activity as compared with that of cisplatin. The toxicity of PC-1 was found quite less than that of cisplatin using MTT, $[^3H]thymidine$ uptake and glucose consumption tests in rabbit proximal tubule cells, human kidney cortical cells and human renal cortical tissues. Based on these results, this novel platinum compound represent a valuable lead in the development of a new anticancer chemotherapeutic agent capable of improving antitumor activity and low toxicity.

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