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The Effect of Acetylcholine on the Intracellular $Ca^{2+}$ Increase of the Mouse Early 2-cell Embryos (생쥐 초기 2-세포 배의 세포내 칼슘 증가에 미치는 Acetylcholine의 영향)

  • Yoon S. Y.;Kang D. W.;Bae I. H.
    • Journal of Embryo Transfer
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    • v.20 no.3
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    • pp.191-200
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    • 2005
  • Many studies have shown that the development of mouse early 2-cell embryos in vitro is related with the intracellular $Ca^{2+}$ changes. In ICR strain mouse, the development of embryos arrests at early 2-cell stage, but the arrested early 2-cell embryos can be rescued by the addition of $Ca^{2+}$-related materials. Acetylcholine (ACh) increases intracellular $Ca^{2+}$ concentration ([$Ca^{2+}$]i) via the mAChR-PLC-IP3 pathway in mouse oocytes. We examined whether ACh rescues 2-cell block in mouse. In early 2-cell embryos, ACh increased [$Ca^{2+}$]i in a dose-dependent manner (p<0.001), and had an effect on rescue of 2-cell block and embryonic development. To identify the signal pathway involved in ACh-induced rescue of 2-cell block, we first applied an agonist of ACh receptor (AChR). Like ACh, carbachol increased intracellular $Ca^{2+}$ concentration ([$Ca^{2+}$]i) and atropine, an antagonist of ACh receptor, blocked the ACh-induced $Ca^{2+}$ increase. In $Ca^{2+}$-free medium, ACh also increased [$Ca^{2+}$]i, indicating that $Ca^{2+}$ increased by ACh is mainly released from the intracellular $Ca^{2+}$ store. The ACh-induced $Ca^{2+}$ increase was blocked by PLC inhibitor (U73122), ryanodine receptor (RyR) antagonist (dantrolene), and CaM KII inhibitor (KN-93), but not by IP3R antagonists (xestospongin C). These results show that ACh increases intracellular $Ca^{2+}$ concentration via mAChR/PLC/RyR, and this contributes to the rescue of 2-cell block.

Modification of Late Radiation Response of Rat Salivary Glands by Pentoxifylline and Diltiazem (쥐의 타액선 방사선조사 후 만성반응에 Pentoxifylline과 Diltiazem이 미치는 영향)

  • Suh, Hyun-Suk;Yang, Kwang-Mo;Kang, Yun-Kyung
    • Radiation Oncology Journal
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    • v.17 no.3
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    • pp.230-237
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    • 1999
  • Purpose : To elucidate the effects of pentoxifylline and diltiazem on the late response of the salivary glands of the rat after irradiation. Materials and Methods : Sixteen Sprague-Dawley rats were divided into 4 groups : (a) irradiation alone (b) irradiation with pentixifylline (PTX) (c) irradiation with diltiazem (DTZ) (d) irradiation with both PTX and DTZ. Irradiation was given in a single fraction of 16 Gy using 4 MV photon energy through an anterior port encompassing the left side of the salivary gland leaving the right side of salivary gland as a control. PTX, 20 mg/kg and/or DTZ, 50 mg/kg were infused intraperitoneally before irradiation, Two rats from each group were sacrificed on the 10th week and the rest was sacrificed on the 16th week after irradiation. Histopathologic examinations were undertaken for each section and the proportion of vacuolated cells out of the total number of cells under light microscopic fields was calculated. The statistical significance in the difference of the proportion of the vacuolated cells among the experimental groups was evaluated by a $x^2$-test. Results : Irradiated salivary glands of the 10th week group revealed markedly increased number of vacuolated cells compared to those of unirradiated control. The proportion of vacuolated cells was significantly reduced in both the PTX group (p value=0.001) and the combined PTX and DTX group compared to those of irradiation alone group. The DTZ alone group did not reveal the significant reduction of vacuolated cells compared to those of irradiation alone group (p value, >0.05). The 16th week groups revealed similar findings to those of the 10th week group, but the degree of chronic inflammatory cell infiltrates and interstitial fibrosis was increased and the number of acinar cells was reduced compared to those of the 10th week group. Conclusions : PTX significantly reduced the late radiation response of salivary glands, but DTZ did not reduce the same degree as PTX did. Taking the positive results of this study into consideration, it seems reasonable to apply PTX into the clinical trial for the head and neck irradiation to reduce the late radiation sequelae of salivary glands in the near future. At the same time the further experiment to clarify the subcellar mechni는 involved in PTX should be preceded.

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Ecological Characteristics of Red Rice (Local Name "SARE," Oryza sativa L.) and Factors Affecting Its Competition with Rice (적미(赤米) (자생도(自生稻), 속명(俗名) "사레")의 생태적(生態的) 특성(特性) 및 벼와의 경합요인(競合要因))

  • Ree, D.W.;Hong, Y.K.;Kim, J.C.;Kim, Y.H.
    • Korean Journal of Weed Science
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    • v.3 no.2
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    • pp.143-150
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    • 1983
  • Red rice (local name "Sore") as a weed has been a serious threat to rice production in direct-seeded rice culture in Ganghwagun, Gyeonggi province. In the Ganghwagun 508 ha out of 1,420 ha in the Samsanmyeon area is infested with red rice. The average lowland rice yield is about 4,300 kg/ha in the Ganghwagun, but the average upland rice yield is about 2,000 kg/ha in infested area. This study was carried out in order to clarify the ecological characteristics of red rice and factors affecting the competitive ability of five red rice varieties, collected from Samsanmyeon in 1981, with rice cultivar. Five varieties-Monggeunsare, Salsare, Ginkaragsare, Galsaegsalsare, Galsaegkaragsare-showed the same morphological characteristics of cultivated Japonica type, Chuncheongbyeo, but red rice tillers more profusely, is taller and lodges more easily than Chucheongbyeo. It shatters easily about 10-15 days after heading date, and at this time the hull is discolored in yellow or dark brown. There are many types of red rice with short or long owns on the spikelet, occasionally with or without own on the spikelet in the same hill, and the grains are short or long. In red reice leaf blast occurs more severely than in cultivated Indica/Japonica type, Teabaegbyeo, particulary serious in Monggeunsare. When red rice invaded in direct-seeded rice, number of panicles of rice became reduced more than other yield components.

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Phytotoxicity Inducing Factors and Its Safening Methods for Benzenesulfonylurea Compound KSC-13906 (Benzenesulfonylurea계 화합물(化合物) KSC-13906의 약해발생요인(藥害發生要因) 및 경감방법(輕減方法))

  • Hwang, I.T.;Choi, J.S.;Hong, K.S.;Yoo, J.H.;Kim, J.S.;Cho, K.Y.
    • Korean Journal of Weed Science
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    • v.18 no.3
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    • pp.225-236
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    • 1998
  • KSC-13906 [Erythro N-{(4,6-dimethoxypyrimidin-2-yl)aminocarbonyl} -2-(2-fluoro-1-hydroxy-n-propyl) benzenesulfonamide, US Patent 5,461,025] was investigated how can control phytotoxicity fluctuation and what a good method apply to new rice herbicide. The growth inhibition was observed when the rice plants was transplanted at a shallow depth(0 - 1cm) and leaching was low(0 - 1cm/ day) from the paddy soil. KSC-13906 appeared to move readily down into the paddy soil with water by 3cm depth in the soil column(${\phi}$ 10cm) filled with loamy sand soil under 3cm/day of leaching condition. Artificial control releasing pattern, designed as treated with KSC-13906 of 9 or 18g ai/ha either at a once or daily treated dividing volume of 1/20, 1/25 and 1/30 of the total volume, increased the safety of KSC-13906 to direct seeded and transplanted rice. The safety of KSC-13906 was also enhanced when KSC-13906 was mixed with dymron. For example, the mixture of KSC-13906 and dymron effectively reduced injury of direct seeded rice plants at 18 and 500g ai/ha, respectively, treated 7 days after transplanting. However, combination of KSC-13906 and several herbicides didn't show any synergistic effetct on herbicidal activity and safening effect on rice. However, the combination of KSC-13906+dymron (9~12+250~500g ai/ha) or KSC-13906+mefenacet+dymron(9+250+250g ai/ha) controlled almost all weeds in paddy field without causing any injury to rice and thus the combination would successfully be used as an oneshot herbicide in rice culture.

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Transformation of Potato using the Phosphinothricin Acetyltransferase Gene as the Selectable Marker Gene (감자의 형질전환을 위한 표지유전자로서 Phosphinothricin Acetyltransferase 유전자의 이용)

  • Jeong, J.H.;Yang, D.C.;Bang, K.S.;Han, S.S.
    • Korean Journal of Weed Science
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    • v.18 no.3
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    • pp.205-213
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    • 1998
  • This experiment was carried out to produce herbicide resistant potatoes hawing only chimeric phosphinothricin acetyltransferase (PAT) genes without using antibiotic selectable marker. The pDY502 vector having only PAT gene was reconstructed for transformation of potato. The reconstructed vector was introduced to Agrobacterium tumefaciens MP90 disarmed, and they were used for potato transformation. Hormonal requirement for plant regeneration from leaves and stem explants of potato was investigated. From this experiment, MS medium treated with IBA 0.1 mg/L + BA 0.5 mg/L was the best for potato regeneration, and the ratio of shoot regeneration was 54% for leaf and 46% for stem in that condition. For transformation, explants of potato leaves and stems were cocultured with A. tumefaciens MP90 containing reconstructed vector harvoring only PAT gene. When the potato explants were placed on various concentrations of bialaphos and all the potato explants were dead on medium with over 5.0mg/L bialaphos. By this selection methods, the explants cocultured with Agrobacterium produced the putative transgenic shoots on medium with 5mg/L bialaphos treatment after 3-4 weeks. Second selection was performed by transferring the shoot tips of putative transgenic to medium containing 20mg/L of bialaphos. The shoot tips grew well on the second selection medium, indicating the production of successful transgenic plants. But normal shoots were dead in same cytotoxic medium. Incorporation of the PAT gene into transgenic potatos were confirmed by PCR analysis of DNA and Southern hybridization. These results show that the PAT gene can serve as a selectable marker and herbicide resistant genes for transformation of potato.

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An Assessment of Post-Injection Transmission Measurement for Attenuation Correction With Rotating Pin Sources in Positron Emission Tomography (양전자방출단층촬영(PET)에서 회전 핀선원과 투과 및 방출 동시 영상 방법을 이용한 감쇠보정 방법 특성에 관한 고찰)

  • Lee, J.R.;Choi, Y.;Lee, K.H.;Kim, S.E.;Chi, D.Y.;Shin, S.A.;Kim, B.T.
    • The Korean Journal of Nuclear Medicine
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    • v.29 no.4
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    • pp.533-540
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    • 1995
  • Attenuation correction is important in producing quantitative positron emission tomography (PET) images. Conventionally, photon attenuation effects are corrected using transmission measurements performed before tracer administration. The pre-injection transmission measurement approach may require a time delay between transmission and emission scans for the tracer studies requiring a long uptake period, about 45 minutes for F-18 deoxyglucose study. The time delay will limit patient throughput and increase the likelihood of patient motion. A technique lot performing simultaneous transmission and emission scans (T+E method) after the tracer injection has been validated. The T+E method substracts the emission counts contaminating the transmission measurements to produce accurate attenuation correction coefficients. This method has been evaluated in experiments using a cylindrical phantom filled with background water (5750 cc) containing $0.4{\mu}Ci/cc$ of F-18 fluoride ion and one insert cylinder (276 cc) containing $4.3{\mu}Ci/cc$. GE $Advance^{TM}$ PET scanner and Ge-68 rotating pin sources for transmission scanning were used for this investigation. Post-injection transmission scan and emission scan were peformed alternatively over time. The error in emission images corrected using post-infection transmission scan to emission images corrected transmission scan was 2.6% at the concentration of $1.0{\mu}Ci/cc$. No obvious differences in image quality and noise were apparent between the two images. The attenuation correction can be accomplished with post-injection transmission measurement using rotating pin sources and this method can significantly shorten the time between transmission and omission scans and thereby reduce the likelihood of patient motion and increase scanning throughput in PET.

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Imaging of Herpes Simplex Virus Type 1 Thymidine Kinase Gene Expression with Radiolabeled 5-(2-iodovinyl)-2'-deoxyuridine (IVDU) in liver by Hydrodynamic-based Procedure (Hydrodynamic-based Procedure를 이용한 간에서의 HSV1-tk 발현 확인을 위한 방사표지 5-(2-iodovinyl)-2'-deoxyuridine (IVDU)의 영상연구)

  • Song, In-Ho;Lee, Tae-Sup;Kang, Joo-Hyun;Lee, Yong-Jin;Kim, Kwang-Il;An, Gwang-Il;Chung, Wee-Sup;Cheon, Gi-Jeong;Choi, Chang-Woon;Lim, Sang-Moo
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.5
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    • pp.468-477
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    • 2009
  • Purpose: Hydrodynamic-based procedure is a simple and effective gene delivery method to lead a high gene expression in liver tissue. Non-invasive imaging reporter gene system has been used widely with herpes simplex virus type 1 thymidine kinase (HSV1-tk) and its various substrates. In the present study, we investigated to image the expression of HSV1-tk gene with 5-(2-iodovinyD-2'-deoxyuridine (IVDU) in mouse liver by the hydrodynamicbased procedure. Materials and Methods: HSV1-tk or enhanced green fluorescence protein (EGFP) encoded plasmid DNA was transferred into the mouse liver by hydrodynaminc injection. At 24 h post-injection, RT-PCR, biodistribution, fluorescence imaging, nuclear imaging and digital wholebody autoradiography (DWBA) were performed to confirm transferred gene expression. Results: In RT-PCR assay using mRNA from the mouse liver, specific bands of HSV1-tk and EGFP gene were observed in HSV1-tk and EGFP expressing plasmid injected mouse, respectively. Higher uptake of radiolabeled IVDU was exhibited in liver of HSV1-tk gene transferred mouse by biodistribution study. In fluorescence imaging, the liver showed specific fluorescence signal in EGFP gene transferred mouse. Gamma-camera image and DWBA results showed that radiolabeled IVDU was accumulated in the liver of HSV1-tk gene transferred mouse. Conclusion: In this study, hydrodynamic-based procedure was effective in liver-specific gene delivery and it could be quantified with molecular imaging methods. Therefore, co-expression of HSV1-tk reporter gene and target gene by hydrodynamic-based procedure is expected to be a useful method for the evaluation of the target gene expression level with radiolabeled IVDU.

Methods for Genetic Parameter Estimations of Carcass Weight, Longissimus Muscle Area and Marbling Score in Korean Cattle (한우의 도체중, 배장근단면적 및 근내지방도의 유전모수 추정방법)

  • Lee, D.H.
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.509-516
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    • 2004
  • This study is to investigate the amount of biased estimates for heritability and genetic correlation according to data structure on marbling scores in Korean cattle. Breeding population with 5 generations were simulated by way of selection for carcass weight, Longissimus muscle area and latent values of marbling scores and random mating. Latent variables of marbling scores were categorized into five by the thresholds of 0, I, 2, and 3 SD(DSI) or seven by the thresholds of -2, -1, 0,1I, 2, and 3 SD(DS2). Variance components and genetic pararneters(Heritabilities and Genetic correlations) were estimated by restricted maximum likelihood on multivariate linear mixed animal models and by Gibbs sampling algorithms on multivariate threshold mixed animal models in DS1 and DS2. Simulation was performed for 10 replicates and averages and empirical standard deviation were calculated. Using REML, heritabilitis of marbling score were under-estimated as 0.315 and 0.462 on DS1 and DS2, respectively, with comparison of the pararneter(0.500). Otherwise, using Gibbs sampling in the multivariate threshold animal models, these estimates did not significantly differ to the parameter. Residual correlations of marbling score to other traits were reduced with comparing the parameters when using REML algorithm with assuming linear and normal distribution. This would be due to loss of information and therefore, reduced variation on marbling score. As concluding, genetic variation of marbling would be well defined if liability concepts were adopted on marbling score and implemented threshold mixed model on genetic parameter estimation in Korean cattle.

Studies on the Nitrate Concentration, Sugar Content and Intake of Four Sorghum X Sudangrass Hybrids Grown under Application of Animal Manure (가축분 시용조건에서 4종 수수 X 수단그라 교잡종의 질산태질소 함량과 당도 및 채식률 비교 연구)

  • Seo, S.;Kim, J.G.;Chung, E.S.;Shin, D.E.;Lim, Y.C.;Yoon, S.H.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.22 no.2
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    • pp.131-136
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    • 2002
  • A field experiment was carried out to compare the nitrate nitrogen concentration, sugar content and animal intake of low $sorghum{\times}sudangrass$ hybrids (Sorghum bicolor (L.) Moench) grown under application of animal manure in 1995. The application amount of animal manure were 40MT in cattle manure, and 30MT/ha in swine and poultry manure. The four NLCF's recommended hybrids used in this study were Pioneer 988, TE-Haygrazer and NC+ 855 (heading type), and Jumbo(headless type). The nitrate nitrogen concentration of forage was 365mg/kg at 50~60cm of plant height, 501mg at 100~120cm, and 502mg at 200~220cm, regardless of animal manure type and cultivar. The nitrate nitrogen concentration of NC+ 855 and P 988 was 519 and 526mg, respectively, and were higher than that(317mg) of Jumbo(P<0.05). The nitrate nioogen concentration of stems was 376mg, and significantly higher than that(135mg) of leaves(P<0.05) regardless of cultivar, and lower parts of stems and leaves were higher than those of upper parts of plants. The sugar contents of four $sorghum{\times}sudangrass$ hybrids were ranged between 3.1 and $3.7^{\circ}$ in cattle manure, 2.9 and $3.3^{\circ}$ in swine manure, and 2.8 and $4.9^{\circ}$ in poultry manure. The sugar content of NC+ 855 was the highest as $3.9^{\circ}$, p 988 was second as $3.4^{\circ}$, and then TE-Haygrazer and Jumbo were lower as $3.2^{\circ}$ and $2.9^{\circ}$, respectively. Animal intake of sorghum ${\times}$ sudangrass hybrids P 988 and NC+ 855 were slightly higher than those of Jumbo and TE-Haygrazer grown under application of cattle manure. Also the average intake of P 988 was a little higher than that of Jumbo. However, there were no significant differences of forage intake among few recommended $sorghum{\times}sudangrass$ hybrids.

MICROLEAKAGE OF MICROFILL AND FLOWABLE COMPOSITE RESINS IN CLASS V CAVITY AFTER LOAD CYCLING (Flowable 및 microfill 복합레진으로 충전된 제 5급와동에서 load cycling 전,후의 미세변연누출 비교)

  • Kang, Suk-Ho;Kim, Oh-Young;Oh, Myung-Hwan;Cho, Byeong-Hoon;Um, Chung-Moon;Kwon, Hyuk-Choon;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.27 no.2
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    • pp.142-149
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    • 2002
  • Low-viscosity composite resins may produce better sealed margins than stiffer compositions (KempScholte and Davidson, 1988: Crim, 1989). Plowable composites have been recommended for use in Class V cavities but it is also controversial because of its high rates of shrinkage. On the other hand, in the study comparing elastic moduli and leakage, the microfill had the least leakage (Rundle et at. 1997) Furthermore, in the 1996 survey of the Reality Editorial Team, microfills were the clear choice for abfraction lesions. The purpose of this study was to evaluate the microleakage of 6 compostite resins (2 hybrids, 2 microfills, and 2 flowable composites) with and without load cycling. Notch-shaped Class V cavities were prepared on buccal surface of 180 extracted human upper premolars on cementum margin. The teeth were randomly divided into non-load cycling group (group 1) and load cycling group (group 2) of 90 teeth each. The experimental teeth of each group were randomly divided into 6 subgroups of 15 samples. All preparations were etched, and Single bond was applied. Preparations were restored with the following materials (n=15) : hybrid composite resin [Z250(3M Dental Products Inc. St. Paul, USA), Denfil(Vericom, Ahnyang, Korea)], microfill [Heliomolar RO(Vivadent, Schaan, Liechtenstein), Micronew(Bisco Inc. Schaumburg, IL, USA)], and flowable composite[AeliteFlo(Bisco Inc. Schaumburg, IL, USA), Revolution(Kerr Corp. Orange, CA, USA)]. Teeth of group 2 were subjected to occlusal load (100N for 50,000 cycles) using chewing simulator(MTS 858 Mini Bionix II system, MTS Systems Corp. Minn. USA). All samples were coated with nail polish 1mm short of the restoration, placed in 2% methylene blue for 24 hours, and sectioned with a diamond wheel. Enamel and dentin/cementum margins were analyzed for microleakage on a sclale of 0 (no leakage) to 3 (3/3 of wall). Results were statistically analyzed by Kruscal-Wallis One way analysis, Mann-Whitney U-test, and Student-Newmann-Keuls method. (p = 0.05) Results : 1. There was significantly less microleage in enamel margins than dentinal margins of all groups (p<0.05) 2. There was no significant between six composite resin in enamel margin of group 1. 3. In dentin margin of group 1, flowable composite had more microleakage than others but not of significant differences. 4. there was no significant difference between six composite resin in enamel margin of group 2. 5. In dentin margin of group 2, the microleakage were R>A =H=M>D>Z. But there was no significant differences. 6. In enamel margins, load cycling did not affect the marginal microleakage in significant degree. 7. In enamel margins, load cycling did affect the marginal microleakage only in Revolution. (p<0.05).