• Journal of Life Science
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• v.17 no.2 s.82
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• pp.198-203
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• 2007

DNA transfection is a powerful tool for studying gene functions. The $Ca^{2+}$-phosphate precipitation remains one of the most popular and cost-effective transfection techniques. Mature neurons are more resistant to transfection than young ones and most other cell types, and easy to die if microenvironment changes. Here, we report a transfection protocol for mature neurons. The critical modifications are inclusion of glial cells in culture and careful control of $Ca^{2+}$-phosphate precipitation under microscope. Cerebral glial cells were grown until ${\sim}70-80%$ confluence in DMEM/10% horse serum, which was thereafter replaced with serum-free Neurobasal/Ara-C, and 319 hippocampal neurons were plated onto the glial layer Formation of fine $DNA/Ca^{2+}$-phosphate precipitates was induced using Clontech $CalPhos^{TM}$ Mammalian Transfection Kit, and the size ($0.5-1\;{\mu}m$ in diameter) and density(about 10 particles/$100\;{\mu}m^2$) were carefully controlled by the time of incubation in the medium. This modified protocol can be reliably applied for transfection of mature neurons that are maintained longer than two weeks in vitro, resulting in 10-15 healthy transfected neurons per a well of 24-well plates. The efficacy of the protocol was verified by punctate expression of $pEGFP-CaMKII{\alpha}$, a synaptic protein, and diffuse expression of pDsRed2. Our protocol provides a reliable method for transfection of mature neurons in vitro.

• Journal of the korean academy of Pediatric Dentistry
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• v.29 no.2
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• pp.139-145
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• 2002

In a primary teeth, dental caries is rapidly advanced the pulp disease, because the primary teeth have the thinner and the weaker enamel layer and the wider pulp chamber than relatively the permanent teeth. And the pulps of primary teeth are exposed during caries removal or even they are exposed by unexpected movement of the children or by trauma. For successive pulp treatment in primary teeth, it is necessary to understand completely about multiple canal morphology, variation of root canal anatomy and specific problem related to root formation and resorption of primary teeth. In upper primary molar, canal configuration of mesial root has the most variation same as upper molar. If not canal treatment is completely, most of all endodontic treatment should be failed. In a clinical case report, upper primary molars existed persistent pain or bleeding during treatment were founded the second MB canal and were performed the endodontic treatment of theirs. As a result, the upper primary molars have no symptom and good prognosis. In the examination of extracted upper primary molar, we found that 8 of 35 teeth(22.8%) in the upper primary first molars and 22 of 33 teeth(66.6%) in the upper primary second molar had the second mesiobuccal canal. It has revealed the high prevalence of two canals in mesiobuccal roots of upper primary molars. The frequency of occurrence of the second mesiobuccal canal must be taken into consideration when endodontic treatment is planned and as a possible cause of otherwise un explained failure.

• Journal of the korean academy of Pediatric Dentistry
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• v.31 no.2
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• pp.228-235
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• 2004

The development and proliferation of the mandibular condyle can be altered by changes in the biomechanical environment of the temporomandibular joint. The biomechanical loads were varied by feeding diets of different consistencies. The purpose of the present study was to determine whether changes of masticatory forces by feeding a soft diet can alter the trabecular bone morphology of the growing mouse mandibular condyle, by means of micro-computed tomography. Thirty-six female, 21 days old, C57BL/6 mice were randomly divided into two groups. Mice in the hard-diet control group were fed standard hard rodent pellets for 8 weeks. The soft-diet group mice were given soft ground diets for 8 weeks and their lower incisors were shortened by cutting with a wire cutter twice a week to reduce incision. After 8 weeks all animals were killed after they were weighed. Following sacrifice, the right mandibular condyle was removed. High spatial resolution tomography was done with a Skyscan Micro-CT 1072. Cross-sections were scanned and three-dimensional images were reconstructed from 2D sections. Morphometric and nonmetric parameters such as bone volume(BV), bone surface(BS), total volume(TV), bone volume fraction(BV/TV), surface to volume ratio(BS/BV), trabecular thickness(Tb. Th.), structure model index(SMI) and degree of anisotropy(DA) were directly determined by means of the software package at the micro-CT system. From directly determined indices the trabecular number(Tb. N.) and trabecular separation(Tb. Sp.) were calculated according to parallel plate model of Parfitt et al.. After micro-tomographic imaging, the samples were decalcified, dehydrated, embedded and sectioned for histological observation. The results were as follow: 1. The bone volume fraction, trabecular thickness(Tb. Th.) and trabecular number(Tb. N.) were significantly decreased in the soft-diet group compared with that of the control group (p<0.05). 2. The trabecular separation(Tb. Sp.) was significantly increased in the soft-diet group(p<0.05). 3. There was no significant differences in the surface to volume ratio(BS/BV), structure model index(SMI) and degree of anisotropy(DA) between the soft-diet group and hard-diet control group (p>0.05). 4. Histological sections showed that the thickness of the proliferative layer and total cartilage thickness were significantly reduced in the soft-diet group.

• Geophysics and Geophysical Exploration
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• v.12 no.1
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• pp.132-142
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• 2009

The Tamar rift valley runs through the City of Launceston, Tasmania. Damage has occurred to city buildings due to earthquake activity in Bass Strait. The presence of the ancient valley, the Tamar valley, in-filled with soft sediments that vary rapidly in thickness from 0 to 250mover a few hundreds metres, is thought to induce a 2D resonance pattern, amplifying the surface motions over the valley and in Launceston. Spatially averaged coherency (SPAC), frequency-wavenumber (FK) and horizontal to vertical spectrum ratio (HVSR) microtremor survey methods are combined to identify and characterise site effects over the Tamar valley. Passive seismic array measurements acquired at seven selected sites were analysed with SPAC to estimate shear wave velocity (slowness) depth profiles. SPAC was then combined with HVSR to improve the resolution of these profiles in the sediments to an approximate depth of 125 m. Results show that sediments thicknesses vary significantly throughout Launceston. The top layer is composed of as much as 20m of very soft Quaternary alluvial sediments with a velocity from 50 m/s to 125 m/s. Shear-wave velocities in the deeper Tertiary sediment fill of the Tamar valley, with thicknesses from 0 to 250m vary from 400 m/s to 750 m/s. Results obtained using SPAC are presented at two selected sites (GUN and KPK) that agree well with dispersion curves interpreted with FK analysis. FK interpretation is, however, limited to a narrower range of frequencies than SPAC and seems to overestimate the shear wave velocity at lower frequencies. Observed HVSR are also compared with the results obtained by SPAC, assuming a layered earth model, and provide additional constraints on the shear wave slowness profiles at these sites. The combined SPAC and HVSR analysis confirms the hypothesis of a layered geology at the GUN site and indicates the presence of a 2D resonance pattern across the Tamar valley at the KPK site.

• Journal of Veterinary Clinics
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• v.20 no.1
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• pp.22-25
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• 2003

The aims of this study were to investigate the efficacy of acupuncture on myoelectrical activity of inflamed or normal colon in the rat, and whether the effect of acupuncture on colonic motility was related to endogenous opioids. Twenty-two male Sprague-Dawley rats were divided into three groups. Experimental groups were normal group (n = 8), colitis group (n = 6) and naloxone group (n = 8). Stainless steel bipolar electrodes were implanted on the serosal layer of the proximal colon of rats. Colitis was induced 7 days after electrode implantation using trinitrobenzene sulphonic acid (TNBS) and ethanol. Electromyograms (EMG) were recorded by using polygraph 11 days after implantation of electrodes. In normal group, normal colonic motility was recorded for 60 min, and then traditional acupuncture at GV-1 was applied for 20 min and EMG was recorded for further 60 min in untreated rats. In colitis group, after recording of basal colonic motility for 60 min, 20 min of acupuncture treatment and further EMG recording was performed for 60 min in TNBS/ethanol treated rats. In naloxone group, following subcutaneous administration of naloxone (3 mg/kg), recording of EMG and acupuncture treatment were performed in TNBS/ethanol treated rats. In the normal group, acupuncture at GV-1 did not induce significant changes in colonic motility. TNBS/ethanol treatment had no significant effect on the frequency of colonic motility. And in colitis group, GV-1 acupuncture significantly decreased colonic motility (p < 0.01). In naloxone group, after injection of naloxone, acupuncture at GV-1 did not change colonic motility in TNBS/ethanol treated rats. On the inflamed colon, naloxone blocked the effect of acupuncture. The present results suggested that endogenous opioids released by acupuncture at GV-1 decrease the motility of inflamed colon in rats, but not normal colon.

• Journal of Veterinary Clinics
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• v.20 no.1
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• pp.12-21
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• 2003

In this study, we evaluated effects of three anastomotic techniques of small intestine on adhesions in the dog. Twenty six healthy mixed dogs were randomly assigned to three groups. Group I(n = 8) was sutured with a simple continuous suture, group II(n = 7) was sutured with a simple interrupted approximating suture and group III(n = 11) was sutured with a single layer continuous Connell suture. On completion of any intestinal anastomosis, a pedicle of greater omentum was wrapped around the suture line in all experimental dogs. One percent sodium carboxymeth-ylcellulose (5ml/kg) was administrated into the abdomen by feeding tube prior to closing the last part of peritoneum in all dogs. Postoperative adhesions were evaluated at 14th day after operation. The adhesions consisted primarily in two dogs in group I, three dogs in group II and group III. There were adhesions between intestinal serosal surfaces in eight dogs in all groups, but there were no intestinal serosa-visceral peritoneum adhesion and intestinal serosa-mesentery adhesion. Mean adhesion scores were less than score 2 in all groups. Between anastomotic site and omental graft, there were 13.13$\pm$4.97 mm (mean$\pm$S.D.) adhesion formation in group I and 17.29$\pm$4.68 mm in group II and 14.64$\pm$3.80mm in group III. A simple continuous suture resulted in the least adhesion formation and a simple interrupted approximating suture resulted in the greatest adhesion formation among the groups. However, there were no significant differences among three suture techniques in the severity of adhesions. Intestinal intussusception only encountered in one dog during the 14 days, the dog operated and survived. Daily monitoring of temperature, activity, appetite, defecation and micturition were done. All of those vital signs were within normal values and there were no obvious differences among the groups. In conclusion, even though there were no significant differences among three groups, a simple continous suture pattern is recommended to prevent adhesions when operating intestinal anastomosis in dogs.

• The Journal of Korean Medicine
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• v.19 no.2
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• pp.125-136
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• 1998

This study is on the etiological factors of the obesity, symptoms of a disease caused by the etiological factors and the method of medical treatment resulted from symptoms of a disease, it is based on Dong-Yi Bo Gam. The conclusion of this study is as follows. 1. The final metabolic of fat is depend on the defensive energy. The shape of fatty layer is changed by the point where the defensive energy is replenish or not. So 'Goin(暠人)' 'Bein(肥人)' and 'Yuckin(肉人)' is made a discriminated and there is defferent between 'Besudaeso(肥瘦大小)' and 'Herlqidaso(血氣多少)'. 2. Etiological factors of obesity is ; essential substance from cereals overcome the primodial energy, spleen and stomach are strong at once, spleen and stomach are weak at once, spleen is disturbed by evil energy, retention of phlegm and fluid, muscle is weak, blood is excess but energy (qi)is weak, hua-sid(華食) eating the sweat food very often, a rich fatty diet. 3. To investigate each syndrome as etiological factons, essential substance from cereals overcome primodial energy is divided into two case: one is the food increase itself because of the weakness of stomach energy (qi), the other is not to digest the food because liver overact spleen by seven emotion. The obesity because of strengthen of the spleen and stomach at once do not feel hungry very well and spleen and stomach do not damaged easily by overacting. The obesity because of weakness of spleen and stomach at once, and disturbace in spleen cause by evil energy, gain the weight even if they eat food a little. Also it is hard to move four limbs. Weakness of muscle is weakness of spleen and stomach. Weakness of muscle is caused by weakness of spleen and stomach. That is the case of faining the weight because of eating food with an empty stomach. The obsity caused by retention of fluid has intestine rumbling and gain the weight suddenly. 4. The method of medical treatment fallowing to the etiological factors: The food is inclined to increase itself caused by weakness of stomach energy, In case of the obseity caused by this phenomenon. We can use Decoction for Reinforcing Middle wanner and Replenishing Qi(補中益氣湯), five tastes il-going-san(左味異功散) and so on. The method of medical treatment on obsity caused by strengthen of spleen and stomach is not referred yet. But we must investigate about using peptic powder(平胃散), ji-chul-huan(枳朮丸) for strengthen of stomach qi (胃氣). If the obesity appear caused by weakness of spleen and stomach and disturbance in spleen as to evil energy, and in the same time stomach qi is weak, we can use li-gong san(異功散). If not only obesity but also indigestion, we can use Decoction for naurishing the stomach and promotion eating(養胃進食湯). If not only obesity but also weakness of spleen and stomach, we can use Decoction for Reinforcing Middle warmer and replenishing Qi(補中益氣湯). The energy being weak, we can use Decoction for six noble drugs(六君子湯). If the obesity appear because of retention of fluid, we can use sin-chul-huan(神朮丸). On the obesit, caused by excess of blood and weakness of qi(血管氣虛), there is not method of mectical treatment yet, but the method of medical treatment that we count of first important maybe to invigorate the spleen and the lung qi and to disperse and recitify the depressed liver energy(疏肝利氣). The method of medical treament about the obesity caused by a rich fatty diet is not mentioned yet, but it is important to improve the lifestyle of food and we think that invigorate the spleen, and the lung-qi(補疏肝氣) and disperse and recitify the depressed liver-energy (??肝利機) amy count of first important. 5.There is useful medicine for weight down for example Herb tea(茶), red bean, wax gourd, etc. 6.The etiological factors of the obesity that revive frequently is weakness of qi(氣虛), from 形成氣衰, deficiency of yang, hasty of qi, damp and so on. The symptoms and sign of a disease that revive frequently to the obesity is appoplexia(中風), leucorrhea, reddish and whitish turbid, feeling of fullness in the upper abdomen, diabetes and so on.

• Journal of the Korean Electrochemical Society
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• v.1 no.1
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• pp.1-7
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• 1998

For the purpose of finding new cathode materials for medium-temperature $(700\~800^{\circ}C)$ solid oxide fuel cells, $Gd_{0.8}Ca_{0.2}Co_{1-x}Fe_xO_3,\;(x=0.0\~0.5)$ are prepared, and their thermal stability and conductivity characteristics are investigated. Also, the cathodic activities are measured after the cathode layer being attached on CGO (cerium-gadolinium oxide) electrolyte disk. The X-ray analyses indicate that the materials prepared by calcining the citrate-gels at $800^{\circ}C$ have the orthorhombic perovskite structure without discernible impurities. The thermal stability of the undoped Co perovskite is so poor that it is decomposed to the individual binary oxide even at $1300^{\circ}C$. But the partially Fe-doped cobaltates exhibit a better thermal stability to retain their structural integrity up to $1400^{\circ}C$. The observation whereby both the undoped and Fe-doped cobaltates melt at ca. $1300^{\circ}C$ leads us to perform the electrode adhesion at <$1300^{\circ}C$. The cathodic activity of $Gd_{0.8}Ca_{0.2}Co_{1-x}Fe_xO_3,\;(x=0.0\~0.5)$, electrodes is superior to $La_{0.9}Sr_{0.1}MnO_3$, among the samples of $x=0.0\~0.5$, the x=0.2 cathode shows the best activity for the oxygen reduction reaction. It is likely that the Fe-doping provides a better thermal stability to the materials but in turn imparts an inferior cathodic activity, such that the optimum trade-off is made at x=0.2 between the two factors. The total electrical conductivity and ion conductivity of $Gd_{0.8}Ca_{0.2}Co_{1-x}Fe_xO_3$, are measured to be 51 S/cm and $6.0\times10^{-4}S/cm\;at\;800^{\circ}C$, respectively. The conductivity values illustrate that the materials are a mixed conductor and the reaction sites can be expanded to the overall electrode surface, thereby providing a better cathodic activity than $La_{0.9}Sr_{0.1}MnO_3$.

• Journal of the Korean Electrochemical Society
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• v.10 no.4
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• pp.283-287
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• 2007

KIER has been developing the anode-supported flat tubular solid oxide fuel cell unit bundle for the intermediate temperature($700{\sim}800^{\circ}C$) operation. Anode-supported flat tubular cells have Ni/YSZ cermet anode support, 8 moi.% $Y_2O_3$ stabilized $ZrO_2(YSZ)$ thin electrolyte, and cathode multi-layer composed of Sr-doped $LaSrMnO_3(LSM)$, LSM-YSZ composite, and $LaSrCoFeO_3(LSCF)$. The prepared anode-supported flat tubular cell was joined with ferritic stainless steel cap by induction brazing process. Current collection for the cathode was achieved by winding Ag wire and $La_{0.6}Sr_{0.4}CoO_3(LSCo)$ paste, while current collection for the anode was achieved by using Ni wire and felt. For making stack, the prepared anode-supported flat tubular cells with effective electrode area of $90\;cm^2$ connected in series with 12 unit bundles, in which unit bundle consists of two cells connected in parallel. The performance of unit bundle in 3% humidified $H_2$ and air at $800^{\circ}C$ shows maximum power density of $0.39\;W/cm^2$ (@ 0.7V). Through these experiments, we obtained basic technology of the anode-supported flat tubular cell and established the proprietary concept of the anode-supported flat tubular cell unit bundle.

• Journal of the Korean Society of Food Science and Nutrition
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• v.24 no.3
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• pp.470-486
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• 1995

Mammary epithelial cells contain a subpopulation of cells with a large proliferativ potential which are responsible for the maintenance of glandular cellularity and are the progenitor cells of mammary cancer. These clonogens give rise to multicellular clonal alveolar or ductal units(AU or DU) on transplantation and hormonal stimulation. To isolate putative mammary clonogens, enzymatically monodispersed rat mammary epithelial cells from organoid cultures and from intact glands are sorted by flow cytometry according to their affinity for FITC labeled peanut lectin(PNA) and PE labeled anti-Thy-1.1 antibody(Thy-1.1) into four subpopulations : cells negative to both PNA and Thy-1.1(B-), PNA+cells, Thy-1.1+cells, and cells positive to both reagents(B+). The in vivo transplantation assays indicate that the clonogenic fractions of PNA+cells from out-growths of organoids in primary cultures for three days in complete hormone medium(CHM) are significantly higher than those of cells from other subpopulations derived from cultrues or from intact glands. Extracellular matrix(ECM) is a complex of several proteins that regulated cell function ; its role in cell growth and differentiation and tissue-specific gene expression. It can act as a positive as well as a negative regulator of cellular differentiation depending on the cell type and the genes studied. Regulation by ECM is closely interrelated with the action of other regulators of cellular function, such as growth factors and hormones. Matrigel supports the growth and development of several different multicellular colonies from mammary organoids and from monodispersed epithelial cells in culture. Several types of colonies are observed including stellate colonies, duct-like structures, two- and three-dimensional web structures, squamous organoids, and lobulo-duct colonies. Organoids have the greatest proliferative potential and formation of multi-cellular structures. Phase contrast micrographs demonstrate extensive intracellular lipid accumulation within the web structures and some of duct-like colonies. At the immunocytochemical and electron micrograph level, casein proteins are predominantly localized near the apical surface of the cells or in the lumen of duct-like or lobulo-duct colonies. Squamous colonies are comprised of several layers of squamous epithelium surrounding keratin pearls as is typical fo squamous metaplasia(SM). All-trans retinoic acid(RA) inhibits the growth of SM. The frequency of lobulo-ductal colony formation increased with the augmentation of RA concentration in these culture conditions. The current study models could provide powerful tools not only for understanding cell growth and differentiation of epithelial cells, but also for the isolation and characterization of mammary clonogenic stem cells.