Purpose: Lingual nerve (LN) damage may be caused by either tumor resection or injury such as wisdom tooth extraction, Although autologous nerve graft is sometimes used to repair the damaged nerve, it has the disadvantage of necessity of another operation for nerve harvesting. Moreover, the results of nerve grafting is not satisfactory. The nerve growth factor (NGF) is well-known to play a critical role in peripheral nerve regeneration and its local delivery to the injured nerve has been continuously tried to enhance nerve regeneration. However, its application has limitations like repeated administration due to short half life of 30 minutes and an in vivo delivery model must allow for direct and local delivery. The aim of this study was to construct a well-functioning $rhNGF-{\beta}$ adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with enhanced and extended secretion of hNGF from the injured nerve by injecting $rhNGF-{\beta}$ gene directly into crush-injured LN in rat model. Materials and Methods: $hNGF-{\beta}$ gene was prepared from fetal brain cDNA library and cloned into E1/E3 deleted adenoviral vector which contains green fluorescence protein (GFP) gene as a reporter. After large scale production and purification of $rhNGF-{\beta}$ adenovirus, transfection efficiency and its expression at various cells (primary cultured Schwann cells, HEK293 cells, Schwann cell lines, NIH3T3 and CRH cells) were evaluated by fluorescent microscopy, RT-PCR, ELISA, immunocytochemistry. Furthermore, the function of rhNGF-beta, which was secreted from various cells infected with $rhNGF-{\beta}$ adenovirus, was evaluated using neuritogenesis of PC-12 cells. For in vivo evaluation of efficacy of $rhNGF-{\beta}$ adenovirus, the LNs of 8-week old rats were exposed and crush-injured with a small hemostat for 10 seconds. After the injury, $rhNGF-{\beta}$ adenovirus($2{\mu}l,\;1.5{\times}10^{11}pfu$) or saline was administered into the crushed site in the experimental (n=24) and the control group (n=24), respectively. Sham operation of another group of rats (n=9) was performed without administration of either saline or adenovirus. The taste recovery and the change of fungiform papilla were studied at 1, 2, 3 and 4 weeks. Each of the 6 animals was tested with different solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) by two-bottle test paradigm and the number of papilla was counted using SEM picture of tongue dorsum. LN was explored at the same interval as taste study and evaluated electro-physiologically (peak voltage and nerve conduction velocity) and histomorphometrically (axon count, myelin thickness). Results: The recombinant adenovirus vector carrying $rhNGF-{\beta}$ was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. GFP expression was observed in 90% of $rhNGF-{\beta}$ adenovirus infected cells compared with uninfected cells. Total mRNA isolated from $rhNGF-{\beta}$ adenovirus infected cells showed strong RT-PCR band, however uninfected or LacZ recombinant adenovirus infected cells did not. NGF quantification by ELISA showed a maximal release of $18865.4{\pm}310.9pg/ml$ NGF at the 4th day and stably continued till 14 days by $rhNGF-{\beta}$ adenovirus infected Schwann cells. PC-12 cells exposed to media with $rhNGF-{\beta}$ adenovirus infected Schwann cell revealed at the same level of neurite-extension as the commercial NGF did. $rhNGF-{\beta}$ adenovirus injected experimental groups in comparison to the control group exhibited different taste preference ratio. Salty, sweet and sour taste preference ratio were significantly different after 2 weeks from the beginning of the experiment, which were similar to the sham group, but not to the control group.
Journal of Korean Academy of Oral and Maxillofacial Radiology
/
v.28
no.1
/
pp.127-143
/
1998
The author evaluated the effects of taxol, a microtubular inhibitor, as a possible radiation sensitizer and the production of prostaglandins on three human cancer cell lines(KB, RPMI-2650 and SW-13) and one murine cell line(L929). Each cell line was divided into four groups (control, taxol only, radiation only and combination of taxol and radiation). The treatment consisted of a single irradiation of 10Gy and graded doses (5, 50, 100, 200, 300, 500 nM) of taxol for a 24-h period. The cytotoxicity of taxol alone was measured at 1 day after(1-day group) and 4 days after(4-day group) the treatment. The survival ratio of cell was analyzed by MTT (3-(4,5-dimethylthiazol-2-yl) -2,5-dimethyl tetrazolium bromide) test. Prostaglandins(PGE2 and PGI2) were measured in the culture medium by a radioimmunoassay. The results obtained were as follows. 1. There was a significantly increased cytotoxicity of KB cells in 4-day group than those in I-day group. There was a high correlation between doses of taxol and cell viability in both groups(l-day group R=0.82741, 4-day group R=0.84655). 2. There was a significantly increased cytotoxicity of RPMI -2650 cells treated with high concentration of taxol in 4-day group than those in I-day group. Also there was a high correlation between doses of taxol and cell viability in 4-day group(R=0.93917). 3. There was a significantly increased cytotoxicity of SW-13 cells treated with high concentration of taxol in 4-day group than those in 1-day group. However no high correlation was observed between doses of taxol and cell viability in both groups(1-day group R=0.46362, 4-day group R=0.65425). 4. There was a significantly increased cytotoxicity of L929 cells treated with low concentration of taxol in 4-day group than those in 1-day group. At the same time, there was a low correlation between doses of taxol and cell viability in both groups(1-day group R=0.34237, 4-day group R=0.23381). 5. In I-day group of L929 cells, higher cytotoxicities were observed in the groups treated with 500 nM taxol than given 10 Gy radiation alone. L929 cells in I-day group alone showed a radiosensitizing effect by taxol.. 6. In addition to L929 cells, all cancer cells treated with a combination of taxol and radiation in 4-day group appeared to have some fragmented nuclei and to float on the medium. In addition, L929 cells appeared to be more confluent. 7. The level of PGE2 production was the highest in the contol KB cells. This appeared to increase in every experimental group of all three cancer cells except L929 cells. There was a significantly increased production of PGE2 in SW -13 cells treated with a combination taxol and radiation compared to the other experimental groups. 8. The level of PGE2 production in the control group of RPMI-Z650 cells was the highest. This appeared to increase in every experimental group of all cells except in SW-13 cells. This also increased significantly in RPMI-2650 cells treated with a combination of taxol and radiation compared to the other experimental groups.
Rail vibration in city zone is becoming a serious environmental problem. In order to make a reduction plan for rail vibration, the research was conducted in which many experiments to measure actual rail vibration along the railroad through the central Deajeon area. A digital vibration level meter was used to measure rail vibration. Vibration levels of Z-axis were measured at every second for the duration of the train passing. The measuring station was placed at every 5m for the distance of 55m. A total of 353 different sets of vibration level were obtained. The signals were processed to get $L_{10}$ value and analyzed in terms of distance, train velocity, and number of trains. As a result, it has been found that rail vibration exceed the allowable vibraton limit of 60 dB, at the point of 25 m far from the railroad center, which is regulated by the las of vibration and noise. Train velocity was found to affect a little for vibration level within the zone. It was also found that a trench installed along a railroad could reduce vibration level up to approximately 10 percent. A model test was conducted to investigate the influence of the location and size of trench, on the transfer of vibration. A heavy steel ball was used to generate vibrations. On the basis obtained from this study, it could be concluded that the application of distance-attenuation and the installment of a trench along railroad could be applied as a reduction plan for rail vibration. Because limitions might exist to depend on the effect of distance attenuation, trenchs excavated along a railroad might be suggested as the most efficient solution to reduce railroad vibration.
Through the screening of marine natural compounds that inhibit cancer cell proliferation, we previously reported that pectenotoxin-2 (PTX-2) isolated from marine sponges exhibits selective cytotoxicity against several cell lines in p53-deficient tumor cells compared to those with functional p53. However, the molecular mechanisms of its anti-proliferative action on malignant cell growth are not completely known. To further explore the mechanisms of its anti-cancer activity and to test whether the status of p53 in liver cancer cells correlates with their chemo-sensitivities to PTX-2, we used two well-known hepatocarcinoma cell lines, p53-deficient Hep3B and p53-wild type HepG2. We have demonstrated that PTX-2 markedly inhibits Hep3B cell growth and induces apoptosis whereas HepG2 cells are much more resistant to PTX-2 suggesting that PTX-2 seems to act by p53-independent cytotoxic mechanism. The apoptosis induced by PTX-2 in Hep3B cells was associated with the modulation of DNA fragmentation factor (DFF) family proteins, up-regulation of pro-apoptotic Bcl-2 family members such as Bax and Bcl-xS and activation of caspases (caspase-3, -8 and -9). Blockade of the caspase-3 activity by caspase-3 inhibitor, z-DEVD-fmk, prevented the PTX-2-induced growth inhibition in Hep3B cells. Moreover, treatment with PTX-2 also induced phosphorylation of AKT and extracellular-signal regulating kinase (ERK), but not c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MARK). Specific inhibitors of PI3K inhibitor (LY294002) and ERK1/2 inhibitor (PD98059) significantly blocks PTX-2-induced-anti-proliferative effects, whereas a JNK inhibitor (SP600125) and a p38 MAPK inhibitor (SB203580) have no significant effects demonstrating that the pro-apoptotic effect of PTX-2 mediated through activation of AKT and ERK signal pathway in Hep3B cells.
The purpose of this study was to evaluate the effects of container types on physiological characteristics of Zelkova serrata, Fraxinus rhynchophylla and Quercus serrata in the container nursery stage. We used 16 container types [4 growing densities (100, 144, 196 and $256\;seedlings/m^2$)${\times}4$ cavity volumes (460, 380, 300 and $220cm^3/cavity$)] and performed two-way ANOVA to test the differences in photosynthesis, photochemical efficiency and chlorophyll content among container types. Also, multiple regression analysis was conducted to correlate container dimensions with photosynthetic rate. Container types had a strong influence on photosynthesis of three species seedlings. Growing densities and cavity volumes had a significant interaction effect on photosynthetic rate, water use efficiency, stomatal conductance and chlorophyll contents except stomatal conductance of Q. serrata. In all three species, however, interactions between the two factors of container type were not found with regard to photochemical efficiency. Growing density was negatively correlated with photosynthetic rate of F rhynchophylla and Q. serrata, while cavity volumes positively affected on those of three species seedlings. The range of optimal container types was determined by multiple regression analysis based on photosynthetic rate. Consequently, optimal growing density and cavity volume of container by each tree species were found to be approximately $160{\sim}210\;seedlings/m^2$ and $430{\sim}460cm^3/cavity$ for Z. serrata, $130{\sim}150\;seedlings/m^2$ and $390{\sim}440cm^3/cavity$ for F. rhynchophylla and $130{\sim}170\;seedlings/m2$ and $420{\sim}460cm^3/cavity$ for Q. serrata, respectively. Application of adequate container will induce higher quality seedling production in nursery stage, which will also increase seedling growth in plantation stage.
Kim, Gi-Ppeum;Ahn, Kyung-Geun;Kim, Gyeong-Ha;Hwang, Young-Sun;Kang, In-Kyu;Choi, Youngmin;Kim, Haeng-Ran;Choung, Myoung-Gun
Horticultural Science & Technology
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v.34
no.1
/
pp.172-182
/
2016
This study was aimed to determine the changes in vitamin $B_5$ and $B_6$ contents compared to fresh materials after parboiling treatment of the main vegetables consumed in Korea. The specificity of accuracy and precision for vitamin $B_5$ and $B_6$ analysis method were validated using high-performance liquid chromatography (HPLC). The recovery rate of standard reference material (SRM) was excellent, and all analysis was under the control line based on the quality control chart for vitamin $B_5$ and $B_6$. The Z-score for vitamin $B_6$ in food analysis performance assessment scheme (FAPAS) proficiency test was -1.0, confirming reliability of analytical performance. The vitamin $B_5$ and $B_6$ contents in a total of 39 fresh materials and parboiled samples were analyzed. The contents of vitamin $B_5$ and $B_6$ ranged from 0.000 to 2.462 and from 0.000 to $0.127mg{\cdot}100g^{-1}$, respectively. The highest contents of vitamin $B_5$ and $B_6$ were $2.462mg{\cdot}100g^{-1}$ in fresh fatsia shoots (stem vegetables), and $0.127mg{\cdot}100g^{-1}$ in fresh spinach beet (leafy vegetables), respectively. Moreover, the vitamin $B_5$ and $B_6$ contents for parboiling treatment in most vegetables were reduced or not detected. In particular, the contents of vitamin $B_5$ in parboiled fatsia shoots and vitamin $B_6$ in parboiled yellow potato and spinach beet were decreased 20- and 4-fold compared with fresh material, respectively. These results can be used as important basic data for utilization and processing of various vegetable crops, information for dietary life, management of school meals, and national health for Koreans.
The aim of this study was to evaluate the effect of cavity shape, bond quality of bonding agent and volume of resin composite on shrinkage stress developed at the cavity floor. This was done by measuring the shear bond strength with respect to iris materials (cavity shape , adhesive-coated dentin as a high C-factor and Teflon-coated metal as a low C-factor), bonding agents (bond quality: $Scotchbond^{TM}$ Multi-purpose and Xeno III) and iris hole diameters (volume; 1mm or 3mm in $diameter{\times}1.5mm$ in thickness). Ninety-six molars were randomly divided into 8 groups ($2{\times}2{\times}2$ experimental setup). In order to simulate a Class I cavity, shear bond strength was measured on the flat occlusal dentin surface with irises. The iris hole was filled with Z250 restorative resin composite in a bulk-filling manner. The data was analyzed using three-way ANOVA and the Tukey test. Fracture mode analysis was also done When the cavity had high C-factor, good bond quality and large volume, the bond strength decreased significantly The volume of resin composite restricted within the well-bonded cavity walls is also be suggested to be included in the concept of C-factor, as well as the cavity shape and bond quality. Since the bond quality and volume can exaggerate the effect of cavity shape on the shrinkage stress developed at the resin-dentin bond, resin composites must be filled in a method, which minimizes the volume that can increase the C-factor.
The purpose of this study was to evaluate the effect of multiple application of all-in-one dentin adhesive system on microtensile bond strength to caries-affected dentin. Twenty one extracted human molars with occlusal caries extending into mid-dentin were prepared by grinding the occlusal surface flat. The carious lesions were excavated with the aid of caries detector dye. The following adhesives were applied to caries-affected dentin according to manufacturer's directions; $Scotchbond^{TM}$ Multi-Purpose in SM group, Adper Prompt $L-Pop^{TM}$ 1 coat in LP1 group, 2 coats in LP2 group, 3 coats in LP3 group, $Xeno^{(R)}$ III 1 coat in XN1 group, 2 coats in XN2 group. and 3 coats in XN3 group. After application of the adhesives, a cylinder of resin-based composite was built up on the occlusal surface. Each tooth was sectioned vertically to obtain the $1{\times}1\;mm^2$ sticks. The microtensile bond strength was determined. Each specimen was observed under SEM to examine the failure mode. Data were analyzed with one-way ANOVA. The results of this study were as follows; 1. The microtensile bond strength values were; SM ($14.38{\pm}2.01$ MPa), LP1 ($9.15{\pm}1.81$ MPa), LP2(14.08{\pm}1.75$ MPa), LP3 ($14.06{\pm}1.45$ MPa). XN1 (13.65{\pm}1.95$ MPa). XN2 ($13.98{\pm}1.60$) MPa, XN3 ($13.88{\pm}1.66$) MPa, LP1 was significantly lower than the other groups in bond strength (p < 0.05). All groups except LP1 were not significantly different in bond strength (p > 0.05). 2. In LP1, there were a higher number of specimens showing adhesive failure. Most specimens of all groups except LP1 showed mixed failure.
Cho, Min Seok;Yang, A-Ram;Hwang, Jaehong;Park, Byung Bae;Park, Gwan Soo
Journal of Korean Society of Forest Science
/
v.110
no.2
/
pp.198-209
/
2021
This study evaluated the effects of the dimensional characteristics of containers on the nitrogen status of Quercus serrata, Fraxinus rhynchophylla, and Zelkova serrata in the container nursery stage. Seedlings were grown using 16 container types [four growing densities (100, 144, 196, and 256 seedlings/m2) × four cavity volumes (220, 300, 380, and 460 cm3/cavity)]. Two-way ANOVA was performed to test the differences in nitrogen concentration and seedling content among container types. Additionally, we performed multiple regression analyses to correlate container dimensions and nitrogen content. Container types had a strong influence on nitrogen concentration and the content of the seedling species, with a significant interaction effect between growing density and cavity volume. Cavity volumes were positively correlated with the nitrogen content of the three seedling species, whereas growing density negatively affected those of F. rhynchophylla. Further, nutrient vector analysis revealed that the seedling nutrient loading capacities of the three species, such as efficiency and accumulation, were altered because of the different fertilization effects by container types. The optimal ranges of container dimension by each tree species, obtained multiple regression analysis with nitrogen content, were found to be approximately 180-210 seedlings/m2 and 410-460 cm3/cavity for Q. serrata, 100-120 seedlings/m2 and 350-420 cm3/cavity for F. rhynchophylla, and 190-220 seedlings/m2 and 380-430 cm3/cavity for Z. serrata. This study suggests that an adequate type of container will improve seedling quality with higher nutrient loading capacity production in nursery stages and increase seedling growth in plantation stages.
Kang, S. M;Kang, N. J;Cho, J. L;Kim, Z. H;Kwon, Y. W
KOREAN JOURNAL OF CROP SCIENCE
/
v.38
no.4
/
pp.350-359
/
1993
The effect of gibberellic acid ($GA_3) and abscisic acid (ABA) on KCl-enhanced proteolysis of senescing leaves of rice(Oryza sativa L. cv. Chilsung) was studied. Emphasis was given to their effects on KCI-enhanced efflux of amino acids and proteinase activity. When treated singly, $GA_3 affected leaf proteolysis little, while ABA increased proteolysis, the rate of amino acid efflux, and ribulose -1,5 -bisphosphate carboxylase / oxygenase (Rubisco)-degrading endoproteinase activity. An additive increase in all three parameters mentioned above was observed when leaves were treated with ABA and KCl. No such an additive effect was found when $GA_3 was treated with KCl. Both $GA_3 and ABA helped to alleviate the KCI-suppressed activity of Rubisco-degrading exoproteinases. The additive increase in proteolysis of rice leaves in the presence of both ABA and KCl could thus be ascribed to a further increase in the efflux of protein hydrolyzates and Rubisco-degrading endoproteinase activity. An increase in proteolysis was accompanied by a decrease in water absorption, and the combined treatment of ABA with KCl resulted in a further reduction of water absorption.
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