• Title/Summary/Keyword: in vitro time

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Comparison of Growth Characteristics, Forage Productivity and Quality between Italian Ryegrass and Synthetics Derived from Interspecific and Intergeneric Hybrids and Ecotypes (이탈리안 라이그라스와 종.속간 잡종 및 생태형 유래 합성종간의 생육특성, 수량성 및 사료가치 비교)

  • Choi, G.J.;Lim, Y.W.;Lim, Y.C.;Sung, B.R.;Kim, M.J.;Kim, K.Y.;Park, G.J.;Kim, S.R.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.22 no.4
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    • pp.265-272
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    • 2002
  • Forage breeding laboratory of National Livestock Research Institute, R.D.A. has made interspecific hybrids of Lolium multiflorum $\times$ L. pratensis and intergeneric hybrids of Lolium $\times$ Festuca since 1984, and has collected ecotypes of Italian ryegrass since 1991. Growth characteristics of these hybrids and ecotypes were researched, and then these clone lines were named. Among these clone lines, the several clones that have polen fertility, high cold-tolerance, and similar heading time were used for making synthetics, Naehan 6, 7, 8, 9, with polycrossing method in 1997. Field experiments were carried out to compare the mophological and agronomical characteristics and forage productivity and quality of the synthetics with those of Italian ryegrass varieties, Barmultra and Hwasan 101. in Suwon and Yonchun from 1999 to 2000. Heading time of the synthetics were 22th to 24th May that belong to late-mature types to be similar to that of Barmultra and Hwasan 101 in Suwon. The synthetics were 101 to 106 c3n in plant length, medium or thick in thickness of stem, dark peen in leaf color, broad and long in flag leaf, strong in lodging resistance, and excellent in regrowth. Winter survivals of the synthetics were no different from that of Barmultra or Hwasan 101 in Suwon, but better than that of Barmultra or Hwasan 101 in Yonchun where was -10 to -12$^{\circ}C$ of minimum average air temperature in January or February. Dry matter(DM) yields of the synthetics were similar to DM 8,238kg per ha of Barmultra in Suwon, but in Yonchun, were more 7 to 13% than DM 7,291kg per ha of Barmultra. Forage qualities, IVDMD, ADF, NDF and TDN of the synthetics were lower than those of Hwasan 101, but higher than those of Barmultra.

Report on the Stability of Freeze-dried Standard Solution (동결건조표준액의 안정성에 관한 보고)

  • Park, Jun Mo;Yoo, Hye Jung;Kim, Han Chul;Han, Geul Soon
    • The Korean Journal of Nuclear Medicine Technology
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    • v.16 no.2
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    • pp.139-148
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    • 2012
  • Purpose : Since standard solution is the one that knows its exact concentration, the curve of the dissolution has been determined according to the amount of the solution, compared to the amount of the unknown sample. Therefore, the antigen that makes up standard materials should be made in a pure form. The configuration of the standard substance solution in the kit we use is a freeze-dried material, or made and comes as a liquid. Lyophilized reference material is used after dissolving in usually D.W. (Distilled Water), and if the antigen to use is too sensitive, reagents should be freeze-dried. Furthermore, when freeze-dried reference has to be frozen again after being dissolved, it should be kept under $-20^{\circ}C$ until the expiration date according to the reports. Since it is not expressed in the experiment if it is safe or stable to reuse the solution which was dissolved a few times, thus, this time it is tested and evaluated that the changes of the standard solution by freezing and melting several times, and its results and the effectiveness of it were compared to the solution which was kept in a fridge. Materials and Methods : Among Vitro diagnostic kits on the market made by radioimmunoassay, parathyroid hormone (PTH), adrenocorticotropic hormone (ACTH), luteinizing hormone (LH) are made of freeze-dried standard solution and all composed of the same Lot.NO. These hormones melted in D.W. and were separated into three groups. In the first group, melting and freezing were repeated, and in the second group, The solution only for one time use was put into a test tube after melting and freeze it. The third group was kept in the refrigerator. This experiment has been conducted from January to February in 2012. January to 2012. PH test was employed because ph is prone to changing depending on the change of protein. Each group of the standard solution, cpm (counter per minute), and the patient relative concentration values were compared by date, and Through the correlation coefficient and Paired t-test, the significant level of each group was analyzed. Results : ACTH, PTH, LH pH values were too subtle denaturation rather than numerical changes in the protein. In addition, when the standard solution of ACTH, PTH, LH was refrigerated, after 3 days and 7 days, there was a significant difference observed between the solution being kept in a refrigerator and a freezer within a significance level. Conclusion : Standard solution should be kept in a freezer, and being kept in a fridge, it is recommended to use the solution as soon as possible.

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Investigation of Helicobacter pylori cagE Locus Diversity on Interleukin-8 Induction in AGS Cell-line (Helicobacter pylori cagE 유전자 다형성이 위상피세포주에서 Interleukin-8 유도능에 미치는 영향)

  • Lee Su-Min;Lee Hak-Sung;Lee In-Suk;Co Yu-Kyung;Han Hae-Won;Oh Jung-Hwan;Park Jae-Myung;Choi Myung-Gyu;Chung In-Sik
    • Journal of Life Science
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    • v.14 no.6 s.67
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    • pp.906-912
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    • 2004
  • Helicobacter pylori infection is highly prevalent, as high as 2/3 of whole population infected, in Korea. H. pylori infection initiates inflammation by induction of interleukin-8 through type IV secretion of CagA. It was recently suggested that induction failure of IL-8 is not associated with defect in cag PAI but associated with cagE locus diversity. This study was designed to investigate ability of 11-8 in-duction according to sequence variation within the cagE gene, cagA TP motifs and vacA m-types in vitro study using AGS cell-line, and to evaluate its association with different clinical outcome. Seventy-four H. pylori stains were isolated from 23 patients with gastric cancer (Ca), 24 subjects with gastritis (G) and 27 patients with duodenal ulcer (Du) in Kangnam St. Mary's Hospital, Seoul, Korea. cagE gene diversity was confirmed by the PCR-RFLP methods with MboI/NlaIII and tyrosine phosphate motifs (TPMs) of cagA was determined TPM-A and C by using DdeI/Tsp5091 restriction enzyme and TPM-B was determend by Real time PCR the method of Owen et al. and IL-8 was measured by ELISA assay. IL-8 activity was positively detected in 59 among 74 strains $(79.7\%)$. IL-8 secretion was significantly increased in MboI A and MboI B type compared to MboI C type and in MboI/NlaIII A-C and B-C type than C-C type. 1L-8 activity was not associated with either the number or composition of cagA tyrosine phosphorylation motifs and vacA m-type. There was no significant difference in IL-8 activity among patient groups. cagE gene diversity is thought to be mainly associated with the induction of IL-8 in H. pylori infection.

Comparative Uptake of Tc-99m Sestamibi and Tc-99m Tetrofosmin in Cancer Cells and Tissue Expressing P-Glycoprotein or Multidrug Resistance Associated Protein (P-Glycoprotein과 Multidrug Resistance Associated Protein을 발현하는 암세포와 종양에서 Tc-99m Sestamibi와 Tc-99m Tetrofosmin의 섭취율 비교)

  • Cho, Jung-Ah;Lee, Jae-Tae;Yoo, Jung-Ah;Seo, Ji-Hyoung;Bae, Jin-Ho;Jeong, Shin-Young;Ahn, Byeong-Cheol;Sohn, Sang-Gyun;Ha, Jeoung-Hee;Lee, Kyu-Bo
    • The Korean Journal of Nuclear Medicine
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    • v.39 no.1
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    • pp.34-43
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    • 2005
  • Purpose: $^{99m}Tc$-sestamibi(MIBI) and $^{99m}Tc$-tetrofosmin have been used as substrates for P-glycoprotein (Pgp) and multidrug resistance associated protein (MRP), which are closely associated with multidrug resistance of the tumors. To understand different handling of radiotracers in cancer cell lines expressing Pgp and MRP, we compared cellular uptakes of $^{99m}Tc$-MIBI and $^{99m}Tc$-tetrofosmin. The effects of cyclosporin A (CsA), well-known multidrug resistant reversing agent, on the uptake of both tracers were also compared. Materials and Methods: HCT15/CL02 human colorectal cancer cells for Pgp expressing cells, and human non-small cell lung cancer A549 cells for MRP expressing cells, were used for in vitro and in vivo studies. RT-PCR, western blot analysis and immunohistochemistry were used for detection of Pgp and MRP. MDR-reversal effect with CsA was evaluated at different drug concentrations after incubation with MIBI or tetrofosmin. Radioactivities of supernatant and pellet were measured with gamma well counter. Tumoral uptake of the tracers were measured from tumor bearing nude mice treated with or without CsA. Results: RT-PCR, western blot analysis of the cells and irnrnunochemical staining revealed selective expression of Pgp and MRP for HCY15/CL02 and A549 cells, respectively. There were no significant difference in cellular uptakes of both tracers in HCT15/CL02 cells, but MIBI uptake was slightly higher than that of tetrofosmin in A549 cells. Co-incubation with CsA resulted in a increase in cellular uptakes of MIBI and tetrofosmin. Uptake of MIBI or tetrofosmin in HCT15/CL02 cells was increased by 10- and 2.4-fold, and by 7.5 and 6.3-fold in A549 cells, respectively. Percentage increase of MIBI was higher than that of tetrofosmin with CsA for both cells (p<0.05). In vivo biodistribution study showed that MIBI (114% at 10 min, 257% at 60 min, 396% at 240 min) and tetrofosmin uptake (110% at 10 min, 205% at 60 min, 410% at 240 min) were progressively increased by the time, up to 240 min with CsA. But increases in tumoral uptake were not significantly different between MIBI and tetrofosmin for both tumors. Conclusion: MIBI seems to be a better tracer than tetrofosmin for evaluating MDR reversal effect of the modulators in vitro, but these differences were not evident in vivo tumoral uptake. Both MIBI and tetrofosmin seem to be suitable tracers for imaging Pgp- and MRP-mediated drug resistance in tumors.

A COMPARATIVE STUDY OF PRESERVING ABILITY OF HUMAN PERIODONTAL LIGAMENT CELLS STORED IN DIFFERENT STORAGE MEDIA (수종의 저장용액에서 치주인대세포의 생존율 비교)

  • Choi, Won-Kyung;Choi, Hyung-Jun;Choi, Byung-Jai;Lee, Jong-Gap
    • Journal of the korean academy of Pediatric Dentistry
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    • v.26 no.2
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    • pp.427-436
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    • 1999
  • Preservation of the remaining periodontal ligament cells on an avulsed tooth is very important to the successful outcome of replantation. HBSS is recommended as the most suitable storage medium for the avulsed tooth that cannot be replanted immediately. But their availability near the site of an accident is doubtful. The purpose of this in vitro study was to compare periodontal ligament cells stored in different storage media obtained easily on the spot. Human periodontal ligament cells were collected from the premolar teeth extracted for orthodontic treatment. Cells were cultured in ${\alpha}-MEM$ culture medium containing 20% FBS, at $37^{\circ}C$ 100% humidity, in a 5% $CO_2$ incubator. Cells were cultured in 96 well culture plate, $5{\times}10^4$ cells per well with ${\alpha}-MEM$ and incubated for 24 hours. After discarding the medium, those cells were cultured in ${\alpha}-MEM$ contained with 10% FBS, pasteurized milk, sterilized saline, unstimulated saliva and bench-dried state at $25^{\circ}C$ room temperature for 30, 60, 90, 120, 180 minutes respectively. And then each group was measured using MTT assay. The results were as follows. 1. Between the group of each time, there was statistically significant difference. Periodontal ligament cells viability was highest in pasteurized milk and was reduced stepwisely in sterilized saline, unstimulated saliva and bench-dried state(p<0.05). 2. between the time of each group, there was statistically significant difference(p<0.05) but was no statistically significant difference at 90-120 minutes in pasteurized milk and at 60-90 minutes and 120-180 minutes in sterilized saline(p>0.05). In conclusion, HBSS as storage medium of an avulsed tooth is not practical on the spot. Insteadily pasteurized milk can be recommended to maintain the periodontal ligament cells viability.

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Comparison of the Uptakes of Tc-99m MIBI and Tc-99m Tetrofosmin in A549, an MRP-expressing Cancer Cell, In Vitro and In Vivo (MRP발현 인체 비소세포 폐암 A549에서 Tc-99m MIBI와 Tc-99m Tetrofosmin섭취의 비교)

  • Yoo, Jeong-Ah;Jeong, Shin-Young;Seo, Myung-Rang;Bae, Jin-Ho;Ahn, Byeong-Cheol;Lee, Kyu-Bo;Choi, Sang-Woon;Lee, Byung-Ho;Lee, Jae-Tae
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.6
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    • pp.382-392
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    • 2003
  • Purpose: Uptakes of Tc-99m MIBI (MIBI) and Tc-99m tetrofosmin (tetrofosmin) in human non-small cell lung cancer A549, multidrug-resistance associated protein (MRP) expressing cell, were investigated in vitro and in vivo. Materials and Methods: Western blot analysis and immunohistochemistry were used for detection of MRP in A549 cells with anti-MRPr1 antibody. Cellular uptakes of two tracers were evaluated at $100{\mu}M$ of verapamil (Vrp), $50{\mu}M$ of cyclosporin A (CsA) and $25{\mu}M$ of butoxysulfoximide (BSO) after incubation with MIBI and tetrofosmin for 30 and 50 min at $37^{\circ}C$, using single cell suspensions at $1{\times}10^6cells/ml$. Radioactivities in supernatants and pellets were measured with gamma well counter. A549 cells were inoculated in each flanks of 24 nude mice. Group 1 (Gr1) and Gr3 mice were treated with only MIBI or tetrofosmin, and Gr2 and Gr4 mice were treated with 70mg/kg of CsA i.p. for 1 hour before injection of 370KBq of MIBI or tetrofosmin. Mice were sacrificed at 10, 60 and 240 min. Radioactivities of organs and tumors were expressed as percentage injected dose per gram of tissue (%ID/gm). Results: Western blot analysis of the A549 cells detected expression of MRPr1 (190 kDa) and immunohistochemical staining of tumor tissue for MRPr1 revealed brownish staining in cell membrane but not P-gp. Upon incubating A549 cells for 60 min with MIBI and tetrofosmin, cellular uptake of MIBI was higher than that of tetrofosmin. Coincubation with modulators resulted in an increase in cellular uptakes of MIBI and tetrofosmin. Percentage increase of MIBI was higher than that of tetrofosmin with Vrp by 623% and 427%, CsA by 753% and 629% and BSO by 219% and 140%, respectively. There was no significant difference in tumoral uptakes of MIBI and tetrofosmin between Gr1 and Gr3. Percentage increases in MIBI (114% at 10 min, 257% at 60 min, 396% at 240 min) and tetrofosmin uptake (110% at 10 min, 205% at 60 min, 410% at 240 min) were progressively higher by the time up to 240 min with CsA. Conclusion: These results indicate that MIBI and tetrofosmin are suitable tracers for imaging MRP-mediated drug resistance in A549 tumors. MIBI may be a better tracer than tetrofosmin for evaluating MRP reversal effect of modulators.

Phytochemical Constituents and Anticancer Activity of Sphagnum palustre Extract (물이끼 추출물의 식물화학적 성분 및 항암활성 연구)

  • Nam, Jung-Hwan;Jeong, Jin-Cheol;Yoon, Young-Ho;Hong, Su-Young;Kim, Su-Jeong;Jin, Yong-Ik;Lee, Ye-Jin;Yoo, Dong-Lim;Lee, Kyung-Tae;Park, Hee-Juhn
    • Korean Journal of Plant Resources
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    • v.24 no.1
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    • pp.40-47
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    • 2011
  • Sphagnum palustre is a semi aquatic moss. S. palustre has been used as Korean traditional medicine to treat cardiac pain and stroke. This study was carried out to analyze phytochemical constituents of S. palustre and investigate the biological activity for the promotion of human health. At first, we isolated seven compounds from the ethanolic extract of this plant, and their structures were characterized by spectroscopic methods. Their structures were characterized to be Coumarin(1), Caffeic acid(2), Quercetin(3), Astragalin(4), Luteolin(5), Chlorogenic acid(6), Rutin(7) were for the first time reported from this source. The ethanol extract from S. palustre which was tested for its anticancer activity against three human tumor cell line by in vitro assay.

Changes in the in vitro Antioxidant and Antithrombosis Activities of Salicornia europaea According to Harvest Time (수확시기에 따른 함초의 항산화 및 항혈전 활성의 변화)

  • Kim, Mi-Sun;Lee, Jung-Nam;Seong, Ha-Jung;Kim, Deuk Hoi;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.26 no.9
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    • pp.1033-1040
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    • 2016
  • In Korea, the aerial parts of the halophyte Salicornia europaea, known as hamcho, are used in salads in April–June and in oriental medicine in September–October In this study, with the aim of developing functional foods to aid blood circulation, hot water extract (HWE) and ethanol extract (EE) were prepared using hamcho harvested from the fields of Shinan, Jeonnam, Korea on 5th April (HWE-04, EE-04), 5th July (HWE-06, EE-06), 5th August (HWE-08, EE-08), 5th September (HWE-09, EE-09), and 5th October (HWE-10, EE-10), and their antioxidant and antithrombosis activities were evaluated. Among the HWEs, HWE-10 showed the highest concentration of total polyphenols and total flavonoids (22.4 and 17.6 mg/ml, respectively), and EE-09 had the highest concentration among the EEs (20.1 and 19.3 mg/ml, respectively). Among the HWEs and EEs, HWE-08 and EE-08 had the highest total sugar and reducing sugar content. In the antioxidation assay, HWE-10 and EE-09 showed strong reducing power, as well as DPPH, ABTS, and nitrite scavenging activities. The calculated RC50s of EE-09 against DPPH, ABTS, and nitrite were 578, 277, and 68.8 μg/ml, respectively. The antithrombosis activity assay revealed that HWE-04, HWE-06, EE-04, and EE-06 had anticoagulation activity against coagulation factors and that HWE-08, HWE-09, EE-08, and EE-09 expressed strong thrombin inhibitory activity, which was comparable to the antithrombosis activity of aspirin. In addition, EE-06 and HWE-08 exhibited strong aggregation inhibitory activities against human platelets. The results suggest that extract from hamcho harvested in particular periods and prepared using a defined solvent has strong potential as a novel food ingredient and an antioxidant and antithrombosis agent.

Study on the Effect of Contact Angles of Elastic Rubber Impression Materials on the Surface of Working Cast (탄성 고무인상재의 접촉각이 작업모형 표면에 미치는 영향)

  • Kim, Joo-Won
    • Journal of dental hygiene science
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    • v.10 no.3
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    • pp.177-183
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    • 2010
  • This study was begun to search effect of contact angles of elastic rubber impression materials on the surface of working cast. Of elastic rubber impression materials with a Type III consistency, such as polysulfide, polyether and addition silicone, we selected one and then measured the contact angle after dripping a distilled water 3.3ml. Then, after pouring a dental anhydrite in three types of impression materials, we prepared a working cast and then examined its surface. Contact angle was measured using a full automatic contact angle measuring system (DM-700, KYOWA, Japan), and the surface of working cast was observed using a field emission scanning electron microscope (JSM-6700F, JEOL Ltd., JAPAN). The following results were obtained: 1) $Mean{\pm}SD$ (SD: standard deviation) of the initial contact angles were $91.3{\pm}20.5^{\circ}$ in the addition silicone materials, $90.0{\pm}2.2^{\circ}$ in the polyethers and $101.5{\pm}2.3^{\circ}$ in the polysulfides. These results indicate that mean values were similar but standard deviations of the three materials showed a great discrepancy. 2) As the time elapsed, addition silicone materials were found to have a contact angle decreased abruptly as compared with the remaining two types. That is, the initial contact angle was $91.3^{\circ}$ and it was abruptly decreased to $29.4^{\circ}$ after 25 seconds. 3) In the polyethers, the initial contact angle was $101.5^{\circ}$ and it was decreased to $90.7^{\circ}$ after 25 seconds. In the polysulfides, however, the initial contact angle was $90.0^{\circ}$ and it was $84.2^{\circ}$ after 25 seconds. This showed almost no changes in the initial contact angles. Moreover, its magnitude was greater than that seen in additional silicones. 4) There were significant differences in the contact angles between the three types of elastic rubber impression materials as the time elapsed (p<0.001). On an observation on the surface of working cast, addition silicone materials were found to have the most dense surface. This was followed by polysulfides and polyethers in a descending order.

INFLUENCE OF LIGHT IRRADIATION OVER SELF-PRIMING ADHESIVE ON DENTIN BONDING (상아질접착제에 대한 광조사가 접착에 미치는 영향)

  • 류현욱;김기옥;김성교
    • Restorative Dentistry and Endodontics
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    • v.26 no.5
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    • pp.409-417
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    • 2001
  • The purpose of this study was to investigate the influence of light irradiation over self-priming adhesive on dentin bonding. After acid etching the exposed dentin, a self-priming adhesive (Prime&Bond$^{\circledR}$NT dental adhesive system Dentsply DeTrey, GmbH, Konstanz, Germany) was applied and light irradiation was done for 20 sec with regular intensity (600 mW/$\textrm{cm}^2$) in group I and for 3 sec with ultra-high intensity (1930 mW/$\textrm{cm}^2$) in group III. No light irradiation was done over self-priming adhesive in groups II and IV. Composite resin was added on the self-priming adhesive and irradiated for 40 sec with regular intensity (600 mW/$\textrm{cm}^2$) in groups I and II and for 3 sec with ultra-high intensity (1930 mW/$\textrm{cm}^2$) in groups III and IV. To see the effect of light curing time on dentin bonding, another 3 group specimens were prepared. Without light-irradiation over self-priming adhesive, added composite resin was irradiated for 3, 6, or 12 sec with ultra-high intensity light. After bonded specimens were stored in 37$^{\circ}C$ distilled water for 24 hours, shear bond strength were measured using a universal testing machine (4202, Instron, Instron Co., U.S.A.) and fractured surfaces were examined under a stereomicroscope (SZ-PT Olympus, Japan). Statistical analysis were done with one-way, two-way ANOVA and chi-square test. The results were as follows : 1. The shear bond strengths from the groups irradiated over self-priming adhesive were significantly higher than those from the groups without irradiation (p<0.05). 2. There was no significant shear bond strength difference between regular intensity light irradiation groups and ultra-high intensity ones (p>0.05). 3. There was no significant shear bond strength difference among various irradiation time groups with ultra-high intensity ones (p>0.05). 4. In stereomicroscopic examination of fractured surfaces, adhesive-cohesive mixed failure mode was mostly seen in all groups, and there was no significant difference in failure mode among groups (p>0.05).

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