• The korean journal of orthodontics
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• v.25 no.4
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• pp.433-445
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• 1995

Tooth movement, the phenomena and mechanisms of which are still controversial, can be considered as part of the result of the inflammatory processes. The purpose of this study was to examine the activity of macrophage and T-cell, playing important roles in the immune reaction, in the periodontal ligament of dog, in which experimental tooth movement was performed. Six one and half year-old dogs, a control and 5 experimentals, were studied. Light force (50-75g) was applied by placing open-coil spring between left mandibular premolars ; heavy force (250-300g), between right mandibular premolars. Experimental dogs were sacrificed at 12 hours, 1, 3, 7 and 14 days since force application, respectively. And the histologic and the immunohistochemical evaluation on the obtained tissue were performed, using $\alpha$-1-antichymotrypsin and CD3 antibodies. The results were as follows : 1. There were more inflammatory cell infiltrations in heavy force group than in light force group until 3 days. But from 7 dsays on, no difference was not observed between groups ; Such an infiltration was more evident at pressure side than at tension side. 2. Osteoclastic activity at pressure side began to be seen in 12 hours, increasing until 7 days. After then it decreased ; Such an activity was more evident in heavy force group than in light force group. 3. Tearing of periodontal ligament and vascular dilatation at tension side began to be seen in 12 hours, increasing until 3 days. After then it decreased ; Such an observation was more evident in heavy force group than in light force group, but there was no difference between groups in 14 days. 4. $\alpha$-1-antichymotrypsin expression in control group was positive, mainly in sulcular epithelium, but negative in periodontal membrane, pulp, bone cells. 5. $\alpha$-1-antichymotrypsin expression in experimental group was more positive in pressure side than in tension side ; The expression was a little more positive in cervical area of tooth until 3 days, but after 7days, it was more positive in apical area. 6. $\alpha$-1-antichymotrypsin expression in light force group began to be observed in 12 hours and reached to the greatest level in 7 days, after which it decreased ; In heavy force group, it was the greatest in 3 days, after which it decreased. 3 Expression in the periodontium was almost negative.

• Applied Microscopy
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• v.30 no.4
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• pp.357-365
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• 2000

Paneth cells have been suggested to contribute to the elimination of excess metals into the intestinal lumen. The purpose of this study wat to investigate the changes of the zinc pools in rats subjected to functional loading with zinc salt by mean of both light and electron microscopical autometallography (AMG). Wistar rats 4 were administrated with zinc chloride (20 mg/kg body weight) intraperitoneally dissolved in 1 ml distilled water. The control group received 1 ml saline IP. After further one hour the animals were transcardially perfused with 0.4% sodium sulphide dissolved in 0.1 M PB fellowed by 3% glutaraldehyde solution for 10 minutes. Pieces of ileum were frozen with solid $CO_2$ and sectioned on a cryostat. The sections $(20{\mu}m)$ were autometallographically developed. Sections selected for EM were reembedded on top of a blank Epon block, from which ultrathin sections (100 nm) were cut. The ultrathin sections were double stained with uranyl acetate (30 min) and lead citrate (5 min), then examined under electron microscope. Studies of comparable sections from control and zinc loaded animals with the AMG selenium method gave quite different results. The control animals demonstrated a weakly positive staining in the cytoplasm of the Paneth cells. In the electron microscope the AMG silver grains were found to be located in the cytoplasm, while the electron dense secretary granules and other cell organelles were void of staining. Few AMG grains were located at the apical surface of the Paneth cells. In sections from zinc loaded rats, the AMG grains were seen in abundance in the lumen of the Lieberkuhn crypts at light microscopic levels. At EM levels the zinc revealing silver grains were located in the cytoplasm as in the controls, but much more AMG grains were shifted into the secretary granules. Furthermore, profound AMG grains were found in the lumen of the crypts and surrounding vessels. And a few grains were seen in the endothelium. The AMG technique demonstrated a pattern of AMG grains in the Paneth cells that strongly suggests a transport of zinc ions through these cells.

• Journal of Sericultural and Entomological Science
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• v.25 no.2
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• pp.1-31
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• 1984

Flacherie, as one of the most prevalent silkworm diseases, causes severe economic damage to sericultural industry and its pathogens have been proved to be flacherie virus (FV) and densonucleosis virus (DNV). Multiplications of the viruses in the larvae of the silkworm, Bombyx mori, were studied by the sucrose density gradient centrifugation and electron microscopy. The quantitative and qualitative changes of nucleic acids and proteins were investigated from the midgut and hemolymph in the silkworm larvae infected separately with FV and DNV. The histopathological changes of epithelial cells of infected midgut also were examined by an electron microscope. 1. Purified fractions of FV or DNV in a sucrose density gradient centrifugation yielded one homogenous and sharp peak without a shoulder, suggesting no heterogenous materials in the preparation. Electron microscopy also revealed that FV and DNV were spherical particles, 27nm and 21nm in diameter, respectively. 2. Silkworm larvae showed a decrease in body weight on the 6th day and in midgut weight on the 3rd day after inoculation with FV or DNV. 3. DNA content was higher in the midgut when infected with FV or DNV, but the hemolymph of the infected larvae showed no difference during first 6 days after inoculation, after which DNA concentration declined rapidly. 4. RNA synthesis of silkworm larvae infected separately with FV and DNV was stimulated in the midgut, but RNA content was reduced in the hemolymph at the early stage of virus multiplication. At the late stage of virus multiplication, however, it was extremely reduced in both midgut and hemolymph. 5. The concentration of protein in the midgut and hemolymph of silkworm larvae infected separately with FV and DNV showed no difference from that of the healthy larvae at the early stage of virus multiplication, but it was significantly reduced at the late stage of virus multiplication. 6. There was no difference in the electrophoretic patterns of RNAs extracted from the midgut of healthy or virus-infected larvae. 7. The electrophoresis of proteins extracted from the midgut infected with FV or DNV, when carried out on the 1st and 5th day after virus inoculation, showed no difference from that of the healthy larvae. But, there was an additional band with medium motility in the proteins on the 8th day after virus inoculation, while a band with low mobility shown in the proteins of healthy larvae disappeared in the infected larvae. However, a band with high mobility in the healthy larvae was separated into two fractions in the infected larvae. 8. The electrophoretic pattern of hemolymph proteins of the silkworm larvae infected separately with FV and DNV was similar to that of the healthy larvae, but the concentration of hemolymph proteins in the infected larvae was lower than that of the healthy larvae at the late stage. 9. Two types of inclusion bodies were shown by the double staining of pyronin-methyl green in the columnar cell of the midgut on the 8th day after FV inoculation. 10. Electron microscopy of the infected midgut revealed that the 'cytoplasmic wall' of the goblet cell thickened on the 5th day after FV inoculation and several types of the cytopathogenic structures, such as virus$.$specific vesicles, virus particles, linear structures, tubular structures, and high electron-dense matrices were observed in the cytoplasm of the goblet cell. The virus particles were also observed in the microvilli and the structures similar to spherical virus particles were observed around the virus-specific vesicles, suggesting the virus assembly in the cytoplasm. 11. Fluorescence micrograph of the infected midgut stained with acridine orange showed that the nucleus, the site of DNV multiplication in the columnar cell, enlarged on the 5th day after virus inoculation. 12. Electron microscopic examination of DNV infected midgut revealed that the nucleolus of the columnar cell was broken into granules and those granules dispersed into apical region of the nucleus on the 5th day after virus inoculation. On the 8th day after inoculation, it was also observed that the nucleus of the columnar cell was full with the high electron-dense virogenic stroma which were similar to virus particles. These facts suggest that the virogenic stroma were the sites of virus assembly in the process of DNV multiplication.

• Horticultural Science & Technology
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• v.28 no.6
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• pp.996-1002
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• 2010

Flow cytometry (FCM) was used to measure the ploidy level of three different sports from 'Campbell Early' ($Vitis$ $labruscana$) grape. Results of the study showed different ploidy levels. FCM analysis for 'Campbell Early' grape which contains 2C DNA diploid cells showed single peak around 35-40 while 'Kyoho' grape with 4C DNA tetraploid cells had a different level of 70-80. However, analysis of the sports displayed a histogram with 2 peaks containing both 2C and 4C nuclei. There was no difference in histograms of 2C DNA flesh and pericarp; on the other hand, 4C DNA flesh type of sports had a different histogram from that of the 2C DNA pericarp. Chromosome numbers of diploid ('Campbell Early'), tetraploid ('Kyoho'), and three sports were counted under the microscope. 'Campbell Early' and 'Kyoho' have 38 and 76 chromosomes, respectively. Three different sports are mixoploids with mixtures of diploid and tetraploid cells. Microscopic observations of shoot apical meristems in sports from 'Campbell Early' grape were carried out to determine the type of plant chimera. 'Campbell Early' grape (diploid) and 'Kyoho' grape (tetraploid) showed that both had 2 tunica layers covering corpus cells, while the three different sports had tunica layers showing mostly oblique division. Most cells from 'Kyoho' grape were larger than 'Campbell Early' grape. Cells from L-2 and L-3 layers of the three sports were similar to 'Kyoho' grape in size, although all cells in L-1 surface layer were uniform in size like 'Campbell Early' grape. Results of FCM analysis indicated that both normal and polyploid cells could be intermixed in sports and could become mixoploidy consisting of diploid and tetraploid. All sports used in the tests were periclinal chimera plants with two distinct L-1 and L-2 cell layers. The result of this study suggests that all three sports which originated from 'Campbell Early' grape might be 2-4-4 type chimera formation.

• Applied Microscopy
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• v.30 no.1
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• pp.45-59
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• 2000

Cis-platin is a widely used anticancer drug against certain solid tumors such as malignant ovarian tumor, malignant carcinoma of head and neck, bladder cancer and cervical cancer of uterus, and its major mechanism of action is inhibition of DNA synthesis of the tumor cell. To investigate the inhibitory effects of cis-platin on the ciliogensis of the ciliated cells in the mucosa of oviduct, the author pursued the alterations of $\alpha-tubulin$, which is the main constituent of the microtubles in cilia, after cis-platin treatment. To eliminate the possible variations due to ovarian cycle, female Spargue-Dawley rats ($150\sim200gm$ in B.W.) were pretreated with estradiol benzoate (20 mg/kg, once a day, for 4 consecutive days). Animals were administrated with cis-platin (6 mg/kg, i.p.) and sacrificed at 1day, 3days, 5days and 7days after treatment, respectively. Immunohistochemistry for $\alpha-tubulin$ using mouse anti-rat $\alpha-tubulin$ monoclonal antibody as primary antibody was done. Immunogold electronmicroscopy for intracellular distributions of $\alpha-tubulin$ was also performed with same primary antibody and Goat anti- mouse IgM which is preconjugated with gold particles of 15 nm as secondary antibody. The results obtained were as follows; 1. Strong immunoreactivity of $\alpha-tubulin$ was observed in ciliated cells of oviducts at 1, 3 and 5 days after estradiol pretreatment. 2. Weak immunoreactivity of $\alpha-tubulin$ was observed in ciliated cells of oviducts at 1 and 3 days after cis-platin treatment but it was recovered to strong immunoreactivity in 5 days 3. In immunogold electronmicroscopy, density of gold particles for $\alpha-tubulin$ reactions was decreased in apical cytoplasm, but few changes were observed in basal body or cilia at 1 and 3 days after cis-platin treatment. From these above results, it is indicated that synthesis of $\alpha-tubulin$ in ciliated cells of rat oviduct is inhibited by cis-platin treatment.

• Journal of Periodontal and Implant Science
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• v.36 no.2
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• pp.555-565
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• 2006

The purpose of the present study is to evaluate the biological stability of the zirconia/alumina composite abutment by histologic and radiographic examination in clinical cases. 17 partially edentulous patients (5 men and 12 women, mean age 47) were treated with 37 implants. The implants were placed following the standard two-stage protocol. After a healing period of 3 to 6 months, zirconia/alumina composite abutments were connected. All radiographs were taken using paralleling technique with individually fabricated impression bite block, following insertion of the prosthesis and at the 3-, 6-, 12 month re-examinations. After processing the obtained images, the osseous level was calculated using the digital image in the mesial and distal aspect in each implant. An ANOVA and t-test were used to test for difference between the baseline and 3-, 6-, 12 months re-examinations, and for difference between maxilla and mandible. Differences at P <0.05 were considered statistically significant. For histologic examination, sample was obtained from the palatal gingiva which implant functioned for 12 months. Sections were examined under a light microscope under various magnifications. Clinically, no abutment fracture or crack as well as periimplantitis was observed during the period of study. The mean bone level reduction(${\pm}standard$ deviation) was 0.34 rom(${\pm}\;0.26$) at 3-months, 0.4 2mm(${\pm}\;0.30$) at 6-months, 0.62 mm(${\pm}\;0.28$) at 12-months respectively. No statistically significant difference was found between baseline and 3-, 6-, 12-months re-examinations (p > 0.05). The mean bone level reduction in maxilla was 0.33(${\pm}0.25$) at 3-months, 0.36(${\pm}0.33$) at 6-months, 0.56(${\pm}0.26$) at 12-months. And the mean bone level reduction in mandible was 0.35(${\pm}0.27$) at 3-months, 0,49(${\pm}0.27$) at 6-months, 0.68(${\pm}0.30$) at 12-months. No statistical difference in bone level reduction between implants placed in the maxilla and mandible. Histologically, the height of the junctional epithelium was about 2.09 mm. And the width was about 0.51 mm. Scattered fibroblasts and inflammatory cells, and dense collagen network with few vascular structures characterized the portion of connective tissue. The inflammatory cell infiltration was observed just beneath the apical end of junctional epithelium and the area of direct in contact with zirconia/alumina abutment. These results suggest the zirconia/alumina composite abutment can be used in variable intraoral condition, in posterior segment as well as anterior segment without adverse effects.

• Journal of Life Science
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• v.31 no.2
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• pp.209-218
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• 2021

This study aimed to evaluate the anti-obesity effects of Cudrania tricuspidata leaf extract in the order of leaf development on the shoot (L0, L1, L2, L3, L4, and L5). The leaves at the apex of a Cudrania tricuspidata shoot were classified as L0; the next leaves of the apex were classified as L1, L2, L3, and L4 from highest to lowest; and the lowest leaf was classified as L5. A series of 70% ethyl alcohol leaf extracts were screened for the inhibitory effects of adipogenesis in 3T3-L1 preadipocytes. We found that the apical leaf extract of Cudrania tricuspidata (CTL0) was the most effective. Next, a study was conducted on the inhibitory action mechanism of CTL0. Treatment with CTL0 significantly suppressed the differentiation of 3T3-L1 preadipocytes in a dose-dependent manner, as confirmed by the decrease in lipid droplet content observed with Oil Red O staining. Treatment with 12.5 ㎍/ml, 25 ㎍/ml, and 50 ㎍/ml of CTL0 significantly reduced the lipid droplet content. Glucose and cellular triglyceride concentrations were reduced in the 3T3-L1 cells on the CTL0-treated medium compared to the differentiation medium (DM control, DMEM + insulin + dexamethasone + rosiglitazone). Compared with DM, CTL0 significantly inhibited the expression of key pro-adipogenic transcription factors, including peroxisome proliferator-activated receptor γ (PPARγ), LPL, A-FABP, and Glut4. These findings show that CTL0 extract has potent anti-obesity effects.

• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.783-794
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• 1996

It has been suggested that ion transport systems are intimately involved in mediating the effects of growth regulatory factors on the growth of a number of different types of animal cells in vivo. The functional importance of the apical membrane $Na^+/H^+$ antiporter in the renal proximal tubule is evidenced by estimates that this transporter mediates the reabsorption of approximately one third of the filtered load of sodium and the bulk of the secretion of hydrogen ions. This study was designed to investigate the pathway utilized by IGF-I in regulating sodium transport in primary cultured renal proximal tubule cells. Results were as follows : 1. $Na^+$ was observed to accumulate in the primary cells as a function of time. Raising the concentration of extracellular NaCl induced an decrease in $Na^+$ uptake compared with control cells in a dose dependent manner. The rate of $Na^+$ uptake into the primary cells was about two times higher in the absence of NaCl($40.11{\pm}1.76pmole\;Na^+/mg\;protein/min$) than in the presence of 140mM NaCl($17.82{\pm}0.94pmole\;Na^+/mg\;protein/min$) at the 30 minute uptake. 2. $Na^+$ uptake was inhibited by IAA($1{\times}10^{-4}M$) or valinomycin($5{\times}10^{-6}M$) treatment($50.51{\pm}4.04$ and $57.65{\pm}2.27$ of that of control, respectively). $Na^+$ uptake by the primary proximal tubule cells was significantly increased by ouabain($5{\times}10^{-5}M$) treatment($140.23{\pm}3.37%$ of that of control). When actinomycin D($1{\times}10^{-7}M$) or cycloheximide($4{\times}10^{-5}M$) was applied, $Na^+$ uptake was decreased to $90.21{\pm}2.39%$ or $89.64{\pm}3.69%$ of control in IGF-I($1{\times}10^{-5}M$) treated cells, respectively. 3. Extracellular cAMP decreased $Na^+$ uptake in a dose-dependent manner($10^{-8}-10^{-4}M$). IBMX($5{\times}10^{-5}M$) also inhibited $Na^+$ uptake. Treatment of cells with pertussis toxin(50pg/ml) or cholera toxin($1{\mu}g/ml$) inhibited $Na^+$ uptake. Extracellular PMA decreased $Na^+$ uptake in a dose-dependent manner(1-100ng/ml). 100 ng/ml PMA concentration significantly inhibited $Na^+$ uptake in IGF-I treated cells. However, staurosporine($1{\times}10^{-7}M$) had no effect on $Na^+$ uptake. When PMA and staurosporine were added together, the inhibition of $Na^+$ uptake was not observed. In conclusion, sodium uptake in primary cultured rabbit renal proximal tubule cells was dependent on membrane potentials and intracellular energy levels. IGF-I stimulates sodium uptake through mechanisms that involve some degree of de novo protein and/or RNA synthesis, and cAMP and/or PKC pathway mediating the action mechanisms of IGF-I.