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Effect of the Isolation Method of Mouse Inner Cell Mass, Types of Feeder Cells and Treatment Time of Mitomycin C on the Formation Rate of ICM Colony (생쥐 내세포괴의 분리방법과 지지세포의 종류와 Mitomycin C 처리 시간이 내세포괴 Colony 형성률에 미치는 영향)

  • Jang, Ho-Jin;Ko, Kyung-Rae;Kim, Mi-Kyung;Na, Yong-Jin;Lee, Kyu-Sup
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.4
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    • pp.265-272
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    • 2006
  • Objective: This study was carried out to evaluate the effect of the isolation methods of inner cell mass from mouse blastocyst, types of feeder cells and treatment time of mitomycin C on the formation rate of ICM colony. Methods: The inner cells were isolated by conventional immunosurgery, partial trophoblast dissection with syringe needles and whole blastocyst co-culture method. Commercially available STO and primary cultured mouse embryonic fibroblast (pMEF) feeder cells were used, and mitomycin C was treated for 1, 2 or 3 hours, respectively. The formation rate of ICM colony was observed after isolation of ICM and culture of ICM on the feeder cells for 7 days. Result: The ICM colony formation rate on STO were significantly higher in partial trophoblast dissection group (58%) than that in immunosurgery (12%) or whole blastocyst culture (16%) group (p<0.05). The formation rate on pMEF feeder layer was higher in partial trophoblast dissection (88%) and whole blastocyst culture (82%) group than that in immunosurgery (16%) group (p<0.05). When mitomycin C treated to pMEF for 2 hours, the formation rate of 88% was significantly higher than those of other conditions. Conclusion: Above results showed that the efficient isolation method of ICM from blastocyst was the partial trophoblast dissection and the appropriate treatment time of mitomycin C was 2 hours. However, the subculture of ICM colony and characterization of stem cells should be carried out to confirm the efficacy of the partial trophoblast dissection method.

Varietal Analysis and Quantification of Resveratrol in Mulberry Fruits (뽕나무 계통별 오디의 레스베라트롤 함량 분석)

  • Kim Hyun-Bok;Kim Jung-Bong;Kim Sun-Lim
    • Journal of Sericultural and Entomological Science
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    • v.47 no.2
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    • pp.51-55
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    • 2005
  • Resveratrol is naturally occurring phytoalexin compounds produced by grape berries, peanuts, and their products in response to stress such as fungal infection, heavy metal ions or UV irradiation. The objective of this study was to develope a reliable high performance liquid chromatographic (HPLC) method for the quantitative determination of trans-resveratrol in mulberry fruit. Samples were extracted in 80% MeOH and filtered with $0.45{\mu}m$ syringe filter. The transresveratrol was separated Waters $C_{18}$ column, using a mobile phase containing 0.025% trifluroacetic acid in 5% acetonitril and 0.035% trifluroacetic acid in 50% acetonitril, detected by photodiode array detector (PDA) at 254 nm and the flow rate was 1ml/min. Under this analytical condition, the mean content of mulberry fruits (fifty varieties) was $777.3{\pm}585.9ppm$. Among the tested samples, 'Mansaengbaekpinosang (II)' was the highest level in 3450.6 ppm. However four accessions including 'Gukbu', 'Sabangso (I)', 'Simseol' and yield mulberry fruit were not able to detected. Eight suitable varieties selected for the production of fruit were 'Jeolgokchosaeng (Chungbuk)' 777.8 ppm, 'Dangsang 7' 771.1 ppm, 'Jangsosang' 133.9 ppm, 'Susungppong' 31.1 ppm, 'Suwonnosang' 639.7 ppm, 'Palcheongsipyung' 1475.9 ppm, 'Kangsun' 864.0 ppm, and 'Jukcheonchosaeng' 1458.5 ppm. 'Daesungppong' which was the first authorized variety for the production of mulberry fruit was 1236.7 ppm. In conclusion, these results suggest that mulberry including fruit and leaf may a good new resource for resveratrol production.

THE EFFECTS OF TRANSFORMING GROWTH FACTOR-$\beta$ ON THE VIABILITY OF HUMAN PERIODONTAL LIGAMENT CELL AND ON THE EXPERIMENTAL TOOTH MOVEMENT IN RAT (Transforming growth factor-$\beta$가 인체 치주 인대세포 활성 및 백서의 실험적 치아 이동에 미치는 영향에 관한 연구)

  • Park, Yun-Kyung;Kim, Sang-Cheol
    • The korean journal of orthodontics
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    • v.28 no.2 s.67
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    • pp.311-327
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    • 1998
  • The purpose of this study was to evaluate the effects of Transforming Growth Factor-${\beta}$ (TGF-${\beta}$) on the viability of human periodontal ligament cells, in-vitro and on the experimental tooth movement in rat, in-vivo. Human periodontal ligaments were cultured from the first premolar tooth extracted for the purpose of the orthodontic treatment. 0.1, 1, 5 and 10ng/m1 of TGF-${\beta}$ was given to the cultured wells, respectively and the viability was evaluated by MTT assay. Twenty Sprague-Dawley rats were divided into 5 experimental groups(4 rats in each) where 100g of force was applied from helical spring across the maxillary incisors. TGF-${\beta}$ was injected via Hamilton syringe into the periodontal ligament at the mesial and the distal surface of a maxillary incisor of 2 rats in each experimental group. Phosphate buffer saline(PBS) was injected in 2 other rats as controls. Experimental groups were sacrificed at 1, 3, 7, 14 and 28 days after force application, respectively. The obtained tissues were evaluated histologically. The obtained results were as follows: 1. The viability of periodontal ligament cells in 0.1ng/ml of TGF-${\beta}$ was not significantly different from that of control at 1-, 2- and 3-day of cultivation. 2. The viability of periodontal ligament cells was significantly increased at 3-day in 1ng/ml or 5ng/ml of TGF-${\beta}$, and at 2-,3-day in 10ng/ml of of TGF-${\beta}$. 3. The zone of hyalinization in periodontal ligament in pressure side was smaller in TGF-${\beta}$ injection group than that in control group at 3-day after the application of experimental force in rat. But no difference was seen after 7-day. 4. Osteoclastic activity and capillary prolieferation in pressure side were greater in TGF-${\beta}$ injection group than that in control group at 3-day to 7-day. 5. Osteoblastic activity and new bone fomation in tension side were greater in TGF-${\beta}$ injection group than that in control group at 3-day to 14-day.

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Development of Rapid Analytical Method of Forbidden Medicines in Dietary Supplements Using LC-ESI-Tandem MS (LC-ESI-tandem MS를 이용한 기능성표방식품 중 부정유해물질 신속검사체계 개발)

  • Kim, Hee-Yun;Jang, Young-Mi;Joo, Hyun-Jin;Jung, Young-Hyun;Lee, Myoung-Sook;Park, Jong-Seok;Lee, Kwang-Ho;Lee, Hwa-Mi
    • Korean Journal of Food Science and Technology
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    • v.39 no.4
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    • pp.372-379
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    • 2007
  • A high-performance liquid chromatography-electrospray ionization (HPLC-ESI) tandem MS was developed for the rapid and simultaneous determination of forbidden medicines in dietary supplements. Thirteen medicinal components such as PDE-5 inhibitors and their analogues, and the newly identified dimethylsildenafil and xanthoanthrafil, were included in this study. After tentative standardization of molecular ions in both polarities using thirteen references on the mass spectrometer, with ESI-continuous infusion via the syringe pump method, the relative intensity of the ions present in the resulting spectra was quantitatively compared. From the results, the ion mode was selected depending on each reference's characteristics. A HPLC method coupled with the ESI mode was developed considering the matrix effect and interference depending on the type of sample. The validation test of the developed method was followed by carrying out precision, accuracy, recovery, sensitivity and linearity, etc. The method showed sufficiently high sensitivity, reproducibility, and specificity, and produced 4 times faster results when compared with the existing HPLC/UV method for the determination of forbidden compounds in dietary supplements.

Harmonization of Hemolysis Index in Clinical Chemistry Laboratory and Its Application as a Result Verification Tool (화학검사 결과의 검증을 위한 용혈 지수 일치화 도구 개발)

  • Pyo, Sang Shin;Nam, Hyun Su;Cha, Young Jong;Lee, Seungkwan;Lee, Hae Kyung
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.350-358
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    • 2017
  • The hemolysis index (HI) is semi-quantitative marker for hemolysis. Because the characteristics of the HI vary from one commercial platform to another, no standardization or harmonization of the HI is currently available. Specimens (N=40) randomly selected from clinical patients were artificially hemolyzed in vitro. The serum of the specimens was then diluted with a 20 mg/dL difference between 0~300 mg/dL based on serum hemoglobin measured using the XE-2100 hematology automation equipment (Sysmex, Japan). Diluted serum was measured using the Hitachi-7600 biochemical automation equipment (Hitachi, Japan) to differentiate between HI and serum hemoglobin. The data showed linearity between HI and serum hemoglobin and that HI 1 contained approximately 20 mg/dL of serum hemoglobin. To determine the blood rejection threshold, the HI was divided into three groups: HI 0~1, HI 4~6, HI 9~15. After another batch of clinical specimens (N=40) was measured using a Hitachi-7600 (Hitachi, Japan), each specimen was moved forward and backward with the piston of the syringe to induce an artificial in vitro hemolysis, then measured again with a Hitachi-7600 (Hitachi, Japan). The percentage difference between the three groups was analyzed by ANOVA or the Kruskal-Wallis test. In the post-test, there were significant differences between the HI 0~1 and the HI 5~6: Glucose, creatinine, total protein, AST, direct bilirubin, uric acid, phosphorus, triglyceride, LDH, CPK, Magnesium, and potassium levels. Because many clinical tests differed significantly, the threshold for hemolysis could be appropriate for HI 5 (serum hemoglobin 100 mg/dL).

Supraclavicular Brachial Plexus block with Arm-Hyperabduction (상지(上肢) 외전위(外轉位)에서 시행(施行)한 쇄골상(鎖骨上) 상완신경총차단(上腕神經叢遮斷))

  • Lim, Keoun;Lim, Hwa-Taek;Kim, Dong-Keoun;Park, Wook;Kim, Sung-Yell;Oh, Hung-Kun
    • The Korean Journal of Pain
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    • v.1 no.2
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    • pp.214-222
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    • 1988
  • With the arm in hyperabduction, we have carried out 525 procedures of supraclavicular brachial plexus block from Aug. 1976 to June 1980, whereas block with the arm in adduction has been customarily performed by other authors. The anesthetic procedure is as follows: 1) The patient lies in the dorsal recumbent position without a pillow under his head or shoulder. His arm is hyperabducted more than a 90 degree angle from his side, and his head is turned to the side opposite from that to be blocked. 2) An "X" is marked at a point 1 cm above the mid clavicle, immediately lateral to the edge of the anterior scalene muscle, and on the palpable portion of the subclavian artery. The area is aseptically prepared and draped. 3) A 22 gauge 3.5cm needle attached to a syringe filled with 2% lidocaine (7~8mg/kg of body weight) and epineprine(1 : 200,000) is inserted caudally toward the second portion of the artery where it crosses the first rib and parallel with the lateral border of the muscle until a paresthesia is obtained. 4) Paresthesia is usually elicited while inserting the needle tip about 1~2 em in depth. If so, the local anesthetic solution is injected after careful aspiration. 5) If no paresthesia is elicited, the needle is withdrawn and redirected in an attempt to elicit paresthesia. 6) If, after several attempts, no paresthesia is obtained, the local anesthetic solution is injected into the perivascular sheath after confirming that the artery is not punctured. 7) Immediately after starting surgery, Valium is injected for sedation by the intravenous route in almost all cases. The age distribution of the cases was from 11 to 80 years. Sex distribution was 476 males and 49 females (Table 1). Operative procedures consisted of 103 open reductions, 114 skin grafts combined with spinal anesthesia in 14, 87 debridements, 75 repairs, i.e. tendon (41), nerve(32), and artery (2), 58 corrections of abnormalities, 27 amputations above the elbow (5), below the elbow (3) and fingers (17), 20 primary closures, 18 incisions and curettages, 2 replantations of cut fingers. respectively (Table 2). Paresthesia was obtained in all cases. Onset of analgesia occured within 5 minutes, starting in the deltoid region in almost all cases. Complete anesthesia of the entire arm appeared within 10 minutes but was delayed 15 to 20 minutes in 5 cases and failed in one case. Thus, our success rate was nearly 100%. The duration of anesthesia after a single injection ranged from $3\frac{1}{2}$ to $4\frac{1}{2}$, hours in 94% of the cases. The operative time ranged from 0.5 to 4 hours in 92.4% of the cases(Table 3). Repeat blocks were carried out in 33 cases when operative times which were more than 4 hours in 22 cases and the others were completed within 4 hours (Table 4). Two patients of the 33 cases, who received microvasular surgery were injected twice with 2% lidocaine 20 ml for a total of $13\frac{1}{2}$ hours. The 157 patients who received surgery on the forearms or hands had pneumatic tourniquets (250 torrs) applied without tourniquet pain. There was no pneumothorax, hematoma or phrenic nerve paralysis in any of the unilateral and 27 bilateral blocks, but there was hoarseness in two, Horner's syndrome in 11 and shivering in 7 cases. No general seizures or other side effects were observed. By 20ml of 60% urcgratin study, we confirm ed the position of the needle tip to be in a safer position when the arm is in hyperabduction than when it is in adduction. And also that the humoral head caused some obstraction of the distal flow of the dye, indicating that less local anesthetic solution would be needed for satisfactory anesthesia. (Fig. 3,4).

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Histochemical Detection of Ionic Zinc in the Rat Olfactory Mucosa: Zinc Selenium Autometallography ($ZnSe^{AMG}$) (랫드 후각점막내 Zinc 이온의 조직화학적 동정)

  • Nam, Dong-Woo;Sun, Yuan-Jie;Kim, Sung-Joo;Kim, Yong-Kuk;Kim, Soo-Jin;Yu, Yun-Cho;Jeong, Young-Gil;Jo, Seung-Mook
    • Applied Microscopy
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    • v.33 no.2
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    • pp.145-154
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    • 2003
  • The present study was designed to demonstrate ionic zinc in the rat nasal mucosa by means of zinc selenium autometallography ($ZnSe^{AMG}$). Rats were given sodium selenide either intraperitoneally (i.p) or intranasally (i.n). Prior to the i.n. administration the rats were anesthetized with pentobarbital sodium (30 mg/kg, i.p.). A thin plastic tube coupled to a Hamilton syringe was then inserted into the right nostril and $10{\mu}l$ of the solution was instilled. For the i.p. administration non-anesthetized rats were given $100{\mu}l$ of the sodium selenide solution (10 mg/kg). Control rats were instilled with saline. After 2 hrs survival, the rats were anaesthetized and transcardially perfused with 3% glutaraldehyde. The olfactory area was removed and put into same fixative. The nose was then sectioned ($30{\mu}m$) horizontally, autometallography (AMG) was performed according to Danscher et al. (1997). After silver enhancement, fine AMG grains were scattered in the whole length of the olfactory epithelium containing olfactory receptor neurons, sustentacular and basal cells. However, much higher concentration of the AMG grains occupied near the surface and in the basal region of the olfactory epithelium. Both groups of i.p. and i.n. administration showed almost same level in the concentration of the AMG grains. In i.n. group, few AMG grains were also found in olfactory nerves of the lamina propria, suggesting zinc transport into the olfactory bulb via olfactory axons. At the electron microscopic level, the AMG grains were most entirely found in the supporting cells of the olfactory epithelium, and they were mostly localized in lysosome-like organelles. The i.n. group showed various signs of tissue damage of the olfactory mucosa, where dense concentration of AMG grains were localized at crystalloid structures. The present study demonstrated dense population of ionic zinc in the rat olfactory epithelium. zinc may play a role in the olfactory functioin and in the pathogenesis of the neurodegerative disorders affecting nose.

Measurement of thermal expansion characteristic of root canal filling materials : Gutta-percha and Resilon (수 종의 근관충전재의 열팽창 특성 측정 : Gutta-percha와 Resilon)

  • Jeon, Kyung-A;Lee, In-Bog;Bae, Kwang-Shik;Lee, Woo-Cheol;Baek, Seung-Ho
    • Restorative Dentistry and Endodontics
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    • v.31 no.5
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    • pp.344-351
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    • 2006
  • The purpose of this study was to evaluate the thermal expansion characteristics of injectable ther-moplasticized gutta-perchas and a Resilon. The materials investigated are Obtura gutta-percha, Diadent gutta-percha, E&Q Gutta-percha Bar and Epiphany (Resilon). The temperature at the heating chamber orifice of an Obtura II syringe and the extruded gutta-percha from the tip of both 23- and 20-gauge needle was determined using a Digital thermometer. A cylindrical ceramic mold was fabricated for thermal expansion test, which was 27 mm long, with an internal bore diameter of 3 mm and an outer diameter of 10 mm. The mold was filled with each experimental material and barrel ends were closed with two ceramic plunger. The samples in ceramic molds were heated in a dilatometer over the temperature range from $25^{\circ}C$ to $75^{\circ}C$. From the change of specimen length as a function of temperature, the coefficients of thermal expansion were deter-mined. There was no statistical difference between four materials in the thermal expansion in the range from $35^{\circ}C$ to $55^{\circ}C$ (p > 0.05). However, Obtura Gutta-percha showed smaller thermal expansion than Diadent and Metadent ones from $35^{\circ}C$ to $75^{\circ}C$ (p < 0.05). The thermal expansion of Epiphany was similar to those of the other gutta-percha groups.

EFFECT OF BENZALKONIUM CHLORIDE ON DENTIN BONDING WITH NTG-GMA/BPDM AND DSDM SYSTEM (Benzalkonium Chloride가 NTG-GMA/BPDM계 및 DSDM계 상아질접착제의 접착성능에 미치는 영향)

  • Shin, Il;Park, Jin-Hoon
    • Restorative Dentistry and Endodontics
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    • v.20 no.2
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    • pp.699-720
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    • 1995
  • This study was conducted to evaluate the effect of benzalkonium chloride solution as a wetting agent instead of water on dentin bonding with NTG-GMA/BPDM system (All-bond 2, Bisco.) and DSDM system (Aelitebond, Bisco.). Benzalkonium chloride solution is a chemical disinfectant widely used in medical and dental clinics for preoperative preparation of skin and mucosa due to its strong effect of cationic surface active detergent. Eighty freshly extracted bovine lower incisor were grinded labially to expose flat dentin surface, and then were acid-etched with 10 % phosphoric acid for 15 second, water-rinsed, and dried for 10 second with air syringe. The specimens were randomly divided into 8 groups of 10 teeth. The specimens of control group were remoistured with water and the specimens of experimental groups were remoistured with 0.1 %, 0.5 %, and 1.0 % benzalkonium chloride solution respectively. And then, the Aelitefil composite resin was bonded to the pretreated surface of the specimens by use of All-bond 2 dentin bonding system or Aelitebond dentin bonding system in equal number of the specimens. The bonded specimens were stored in $37^{\circ}C$ distilled water for 24 hours, then the tensile bond strength was measured, the mode of failure was observed, the fractured dentin surface were examined under scanning electron microscopy, and FT-IR spectroscopy was taken for the purpose of investigating the changes of the dentin surface pretreated with benzal konium chloride solution followed by each primer of the dentin bonding systems. The results were as follows : In the group of bonding with NTG-GMA/BPDM dentin bonding agent(All-bond 2), higher tensile bond strength was only seen in the experimental group remoistured with 0.1 % benzal konium chloride solution than that in water-remoistured control group(p<0.05). In the group of bonding with DSDM dentin bonding agent (Aelitebond), no significant differences were seen between the control and each one of the experimental group(p<0.05). Higher tensile bond strength were seen in NTG-GMAIBPDM dentin bonding agent group than in DSDM dentin bonding agent group regardless of remoistur ization with benzal konium chloride solution. On the examination of failure mode, cohesive and mixed failure were predominantly seen in the group of bonding with NTG-GMAIBPDM dentin bonding agent, while adhesive failure was predominantly seen in the group of bonding with DSDM dentin bonding agent. On SEM examination of fractured surfaces, no differences of findings of primed dentin surface between the groups with and without remoisturization with benzal konium chloride solution. FT-IR spectroscopy taken from the control and the experimental group reve::.led that some higher absorbance derived from the primers binding to dentin surface was seen at the group pretreated with 0.1 % benzal konium chloride solution than at the control group of remoisturizing with water.

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SLUMPING TENDENCY AND RHEOLOGICAL PROPERTY OF FLOWABLE COMPOSITES (Flowable 복합레진의 slumping 경향과 유변학적 성질)

  • Lee, In-Bog;Min, Sun-Hong;Kim, Sun-Young;Cho, Byung-Hoon;Back, Seung-Ho
    • Restorative Dentistry and Endodontics
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    • v.34 no.2
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    • pp.130-136
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    • 2009
  • The aim of this study was to develop a method for measuring the slumping resistance of flowable resin composites and to evaluate the efficacy using rheological methodology. Five commercial flowable composites (Aelitefil flow:AF, Filtek flow:FF, DenFil flow:DF, Tetric flow:TF and Revolution:RV) were used. Same volume of composites in a syringe was extruded on a glass slide using a custom-made loading device. The resin composites were allowed to slump for 10 seconds at $25^{\circ}C$ and light cured. The aspect ratio (height/diameter) of cone or dome shaped specimen was measured for estimating the slumping tendency of composites. The complex viscosity of each composite was measured by a dynamic oscillatory shear test as a function of angular frequency using a rheometer. To compare the slumping tendency of composites, one way-ANOVA and Turkey's post hoc test was performed for the aspect ratio at 95% confidence level. Regression analysis was performed to investigate the relationship between the complex viscosity and the aspect ratio. The results were as follows. 1. Slumping tendency based on the aspect ratio varied among the five materials (AF