• Journal of Nutrition and Health
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• v.57 no.1
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• pp.53-64
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• 2024

Purpose: Mitochondria play a crucial role in preserving skeletal muscle mass, and damage to mitochondria leads to muscle mass loss. This study investigated the effects of oxypeucedanin hydrate, a furanocoumarin isolated from Angelica dahurica radix, on myogenesis and mitochondrial function in vitro and in zebrafish models. Methods: C2C12 myotubes cultured in media containing 0.1, 1, 10, or 100 ng/mL oxypeucedanin hydrate were immunostained with myosin heavy chain (MHC), and then multinucleated MHC-positive cells were counted. The expressions of markers related to muscle differentiation, muscle protein degradation, and mitochondrial function were determined by quantitative reverse transcription polymerase chain reaction. To investigate the effects of oxypeucedanin hydrate on mitochondrial dysfunction, Tg(Xla.Eef1a1:mito-EGFP) zebrafish embryos were treated with 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI) with or without oxypeucedanin hydrate and analyzed for mito-EGFP intensity and mitochondrial length. Results: Oxypeucedanin hydrate significantly increased MHC-positive multinucleated myotubes (≥ 3 nuclei) and increased the expression of the myogenic marker myosin heavy chain 4. However, it decreased the expressions of muscle-specific RING finger protein 1 and muscle atrophy f-box (markers of muscle protein degradation). Furthermore, oxypeucedanin hydrate enhanced the expressions of markers of mitochondrial biogenesis (peroxisome proliferator-activated receptor-gamma coactivator 1 alpha, transcription factor a mitochondrial, succinate dehydrogenase complex flavoprotein subunit A, and cytochrome c oxidase subunit 1) and mitochondrial fusion (optic atrophy 1). However, it reduced the expression of dynamin-related protein 1 (a mitochondrial fission regulator). Consistently, oxypeucedanin hydrate reduced FOLFIRI-induced mitochondrial dysfunction in the skeletal muscles of zebrafish embryos. Conclusion: The study indicates that oxypeucedanin hydrate promotes myogenesis by improving mitochondrial function, and thus, suggests oxypeucedanin hydrate has potential use as a nutritional supplement that improves muscle mass and function.

• Parasites, Hosts and Diseases
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• v.24 no.2
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• pp.121-136
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• 1986

Recently there have been some reports on human infections of Echinostoma hortense in Korea. It was found that a few species of freshwater fishes were playing the role of the second intermediate host of E. hortense. However, molluscan intermediate host has not been identified yet in Korea. The present study aimed to establish the life cycle of E. hortense in laboratory. Experimental studies such as egg production from the rat, development of the eggs in vitro, exposure of miracidia to freshwater snails, shedding pattern of cercariae from infected snails, morphology of cercariae, cercarial infection to the second intermediate host and infection of metacercariae to the definitive hosts were done. In addition, epidemiological surveys on the infection status in inhabitants and house rats, and on the natural infection of larval echinostomes in the snails and fishes were carried out along the South Hangang-river. The results obtained were as follows: 1. The eggs deposited from adults in physiological saline were cultivated at room temperature($20{\sim}24^{\circ}C$). The miracidia were firstly observed on 8 days after cultivation, and 85.5% of the eggs contained the mature miracidia on 11 days after cultivation. More than 90% formed the miracidia when cultivated at temperature $22{\sim}27^{\circ}C$. Hatching of the miracidia began on 12 days after cultivation and continued for a week. The size of the miracidia was $103.0{{\times}}51.4{\mu}m$ in average. The motility of the miracidia were active up to 8 hours after shedding, but they were all dead within 10 hours after shedding. 2. A freshwater snail, Radix auricularia coreana was cultivated in aquaria. A hatched $F_1$ snails from the egg masses were exposed to 20 miracidia respectively. Escape of cercariae started on 15 days after infecton. Radix auricularia coreana was experimentally identified as the first intermediate host of E. hortense in Korea. 3. Cercarial shedding started on $15{\sim}20$ days after infection by snail, continued for about 10 days (8.8 days in average). Infected snails were dead within 32 days after the miracidial infection. About 1,335 cercariae($328{\sim}1,994$) per snail were shed in its life, and 119 cercariae in average per snail per day were shed. The cercariae were motile for more than 24 hours, and then squirming at the bottom until death. The body and tail sizes of cercariae were $356{\times}186{\mu}m$ and $510{\times}68{\mu}m$ in average, respectively. The rediae parasitized in the snail hosts were found mainly around the pericardial regions, and their size was $1,575{\times}258{\mu}m$ in average. The numbers of developing cercariae in a mature redia were 14 in average ($7{\sim}20$ in range). The numbers of rediae in a snail were 102 in average on 15 days after miracidial infection and 221 in average on 28 days. 4. Three uninfected Misgurnus anguillicaudatus, less than 6.5cm long were used in for the cercarial infection. They were all exposed with 755 cercariae, and examined at 5-day intervals starting from 10 days after infection. All the fishes were infected with metacercariae of E. hortense and a total of 275 was found infected(36.4%). The metacercariae were fed to rats and the adult worms were obtained on 15 days after infection. 5. The infected rats began to deposit the eggs on 11 days after infection. The number of eggs deposited per day per worm (EPD/worm) was $400{\sim}500$ on 3 weeks after infection and was increased to $1,000{\sim}1,500$ on 4 to 17 weeks, then decreased to 800 on 21 weeks after infection. 6. A total of 745 stool specimens collected from 576 male and 169 female residents of 8 different villages along South Hangang basin was examined. Out of 745 specimens, the eggs of Echinostoma sp. were found in 2 cases (0.3%). Of 34 house rats one showed egg-positive (2.9%). 7. Total 971 Radix auricularia coreana collected from 7 sampling stations were examined for shedding of cercariae. Three snails (0.3%) shed the cercariae of E. hortense. A total of 119 out of 542 freshwater fishes (22.0%) had the metacercariae of E. hortense. The fishes parasitized with the metacercariae were 4 out of 14 examined species. The infection rates of 4 species were 34.1% (106 out of 311) in Misgurnus anguillicaudatus, 30.4% (7 out of 23) in Misgurnus mizolepis, 4.3% (2 out of 46) in Moroco oxycephalus and 22.2% (4 out of 18) in Odontobutis obscura interrupta. In summarizing the above results, the first intermediate host of E. hortense was found as Radix auricularia coreana in Korea. Also, it took about 46 days for the shortest completion of a life cycle of E. hortense in summer; that is, 10 days for miracidial development in eggs, 15 days for cercarial development in the snail, about 10 days for metacercarial development in the second intermediate hosts, and 11 days for the maturation as the adults in the definitive hosts. The natural infection rates of E. hortense in the intermediate hosts were relatively high but those in the definitive hosts were low in the middle areas of South Hangang basin.

• Restorative Dentistry and Endodontics
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• v.33 no.2
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• pp.148-161
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• 2008

The purpose of this study was to evaluate the effect of chlorhexidine (CHX) on microtensile bond strength (${\mu}TBS$) of dentin bonding systems. Dentin collagenolytic and gelatinolytic activities can be suppressed by protease inhibitors, indicating that MMPs (Matrix metalloproteinases) inhibition could be beneficial in the preservation of hybrid layers. Chlorhexidine (CHX) is known as an inhibitor of MMPs activity in vitro. The experiment was proceeded as follows: At first, flat occlusal surfaces were prepared on mid-coronal dentin of extracted third molars. GI (Glass Ionomer) group was treated with dentin conditioner, and then, applied with 2 % CHX. Both SM (Scotchbond Multipurpose) and SB (Single Bond) group were applied with CHX after acid-etched with 37% phosphoric acid. TS (Clearfil Tri-S) group was applied with CHX, and then, with adhesives. Hybrid composite Z-250 and resin-modified glass ionomer Fuji-II LC was built up on experimental dentin surfaces. Half of them were subjected to 10,000 thermocycle, while the others were tested immediately. With the resulting data, statistically two-way ANOVA was performed to assess the ${\mu}TBS$ before and after thermo cycling and the effect of CHX. All statistical tests were carried out at the 95 % level of confidence. The failure mode of the testing samples was observed under a scanning electron microscopy (SEM). Within limited results, the results of this study were as follows; 1. In all experimental groups applied with 2 % chlorhexidine, the microtensile bond strength increased, and thermo cycling decreased the micro tensile bond strength (P > 0.05). 2. Compared to the thermocycling groups without chlorhexidine, those with both thermocycling and chlorhexidine showed higher microtensile bond strength, and there was significant difference especially in GI and TS groups. 3. SEM analysis of failure mode distribution revealed the adhesive failure at hybrid layer in most of the specimen. and the shift of the failure site from bottom to top of the hybrid layer with chlorhexidine groups. 2 % chlorhexidine application after acid-etching proved to preserve the durability of the hybrid layer and microtensile bond strength of dentin bonding systems.

• The Journal of Korean Academy of Prosthodontics
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• v.47 no.1
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• pp.61-69
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• 2009

Statement of problem: Porcelain veneers have become a popular treatment modality for aesthetic anterior prosthesis. Fitting porcelain veneers in the mouth usually involve a try-in appointment, which frequently results in salivary contamination of fitting surfaces. Purpose: An in vitro study was carried out to investigate the effect of silane treatment timing and saliva contamination on the resin bond strength to porcelain veneer surface. Material and methods: Cylindrical test specimens (n=360) and rectangular test specimens (n=5) were prepared for shear bond test and contact angle analysis. Whole cylindrical specimens divided into 20 groups, each of which received a different surface treatment and/or storage condition. The composite resin cement stubs were light-polymerized onto porcelain adherends. The shear bond strengths of cemented stubs were measured after dry storage and thermocycling (3,000 cycles) between 5 and $55^{\circ}C$. The silane and their reactions were chemically monitored by using Fourier Transform Infrared Spectroscopy analysis (FTIR) and contact angle analysis. One-way analysis of variance (ANOVA) and Dunnett's multiple comparison were used to analyze the data. Results: FT-IR analysis showed that salivary contamination and silane treatment timing did not affect the surface interactions of silane. Observed water contact angles were lower on the saliva contaminated porcelain surface and the addition of 37% phosphoric acid for 20 seconds on saliva contaminated porcelain increased the degree of contact angle. Silane applied to the porcelain, a few days before cementation, resulted in increasing the bond strength after thermocycling. Conclusion: Within the limitation of this study, it can be concluded that it would be better to protect porcelain prosthesis before saliva contamination with silane treatment and to clean the contaminated surface by use of phosphoric acid.

• Asian Journal of Turfgrass Science
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• v.20 no.1
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• pp.65-76
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• 2006

Scz1, an isolate of Sclerotinia homoeocarpa, was recently reported as a novel pathogen responsible for dollar spot disease in Zoysiagrass, a warm season turfgrass. Scz1 possessed different characteristics on mycelial pigment, mycelial affinity and host pathogenecity compared to those of Scb1, a typical isolate, obtained from creeping bentgrass, a cool season turfgrass. In this study, only three isolates, Scz1, Scz2(another analogous isolate of Sclerotinia homoeocarpa from zoysiagrass), and Scb1, were examined at the molecular level using the internal transcribed spacer(ITS) and random amplified polymorphic DNA(RAPD) assays to verify their identification and genetic variation. As a result of ITS assay, partial ITS sequences of three isolates showed 94-97% similarity with a standardized ITS sequence of S. homoeocarpa registered on BLAST. In the analysis of RAPD, range value through similarity matrix was 0.167 between Scz1 and Scb1, 0.139 between Scz2 and Scb1, and 0.713 between Scz1 and Scz2, respectively. Furthermore, tendegram analysis indicated that Scz1 and Scz2, unlike Scb1, were clustered together as accompanying a high genetic similarity. In in vitro fungicide bioassay, $EC_{50}$ value representing the sensitivity degree to propiconazole, a well-known fungicide for dollar spot disease, was 0.012 ${\mu}g/ml$ for Sczl, 0.003 ${\mu}g/ml$ for Scz2, and 0.030 ${\mu}g/ml$ for Scb1. From all data taken, we concluded that both Scz1 and Scz2 belonged to one group of S. homoeocarpa, since they exhibit the same host range and high level of genetic similarity, whereas their chemical competences to a fungicide were different. This study would provide further approach for assessing genetic diversity of S. homoeocarpa isolates as well as characterizing individual isolate against chemical exposure.

• Korean Journal of Animal Reproduction
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• v.22 no.1
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• pp.1-9
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• 1998

This study was carried out to confirm whether the developmental capacity of bovine mature oocytes frozen ultra-rapidly using electron microscopic(EM) grids and EFS30 can be obtained, and whether the cryoprotectants and the freezing method used in this study effect detrimentally to the bovine oocytes by indirect immunocytochemistry. As freezing solution, we used EFS30 which consisted of 30% ethylene glycol, 0.5 M sucrose, 18% ficoll and 10% FBS added in D-PBS. The results obtained in this experiment were summarized as follows: When the effects of cryoprotectant and freezing procedure on the microtuble, micrfilament and chromatin morphology of oocytes were evaluated using indirect immunocytochemistry, the results of freezing as well as exposure group were not different with that of the control oocytes. When the fertilization abnormality after ultrarapid freezing of bovine mature oocytes was examined by Hoechst staining, the rates of total penetration(96.7, 9.0%), normal two pronuclei formation(74.6, 68.9%) and mean number of sperm / oocyte(1.50, 1.44) were not different between control and freezing group. In addition, when the developmental capacity of frozen-thawed of oocytes(85.5%) was survived, 74.5% of them were cleaved and 31.4% of cleaved embryos were developed to blastocyst. These data were similar to those of the control(76.0%, 34.6%) and exposure(74.5%, 33.0%) except survival rates. Also, when the total cell number of blastocysts produced from the each treatment at day after IVF was examined by hoechst staining, there were not different among groups. There results demonstrate that developmental capacity of frozen-thawed bovine mature oocytes can be successfully obtained by ultra-rapid freezing method using EM grid and EFS30 solution.

• The Korean Journal of Pesticide Science
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• v.16 no.3
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• pp.261-266
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• 2012

It is common to use many experiment animals to evaluate the toxicity of chemicals including pesticides. For protecting animal, the concepts of 3R (Reduction, Replacement, Refinement) were introduced and in vitro alternatives methods actively have been developed all over the world. Many experimental animals for toxicological tests have been used, so that it is important to establish the alternative methods. In this study, the alternative method using reconstituted human skin model (Keraskin$^{TM}$) was conducted for classification of skin irritation on pesticides. Sixteen formulations selected on the basis of the degree of irritation were treated by Keraskin$^{TM}$ test. The percent of cell viability was measured into the culture medium collected after treatment of the pesticides for 24-72 hrs. The skin irritations of formulations were evaluated by the cell viability. In this study, The 4 formulations with mild irritation in rabbits were evaluated as nonirritant, the 6 formulations with moderate and severe irritation were evaluated as irritant in human skin model test. We suggest that the alternative test using Keraskin$^{TM}$ model could be used as toxicity evaluation for primary irritation index (P.I.I.) score of greater than or equal to 2.1 of pesticides. The further studies should be required to apply for hazardous assessment of pesticides on alternative skin irritation methods because of the interindividual variability of the sensitivity of skin irritation on pesticides.

• Journal of the korean academy of Pediatric Dentistry
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• v.34 no.1
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• pp.1-12
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• 2007

The purpose of this study was to evaluate the effect that various concentration and application time of hydrogen peroxide had on tooth whitening and physical properties. The hydroxyapatite (HA) discs of $12mm({\Phi}){\times}1.2mm(t)$ in dimensions were made by compression $(100kg/cm^2)$ and sintering (at $1350^{\circ}C$ for 2 hours) All specimens were polished sequentially with '240 through '2000 emery paper and one side of each specimen was polished finally with $0.3{\mu}m$ alumina paste. The discs were placed in sterile whole stimulated saliva overnight at $37^{\circ}C$ in order to form an in vitro pellicle layer. Then the discs were rinsed with distilled water and soaked into staining broth at $37^{\circ}C$ for 7 days. These stained specimens were bleached with hydrogen peroxide according to the change of concentration $(3{\sim}30%)$ and application time ($3{\sim}10$ days). The specimens were analyzed with a spectrophotometer, X-ray diffractometer (XRD), scanning electron microscope (SEM), surface roughness tester, microhardness tester and biaxial flexural strength. The results of present study can be summarized as follows : 1. The bleaching effect was increased with the increased concentration and the extended application time of hydrogen peroxide. 2. The surface roughness was significantly increased from the specimen bleached with 15% hydrogen peroxide for 10 days and with 30% for 7 and 10 days respectively (p<0.05). 3. The changes of crystal phase observed by XRD between before and after bleaching weren't shown of any difference, but microporous structure of surface observed by SEM was shown of increase with the increased concentration and the extended application. 4. The biaxial flexural strength was significantly decreased from bleaching of specimen with 30% hydrogen peroxide for 7 and 10 days respectively (p<0.05) 5. Microhardness was significantly decreased from bleaching with 15% hydrogen peroxide for 10 days and with 30% for 3, 7 and 10 days respectively (p<0.05). Although the tooth bleaching effect was greater when the high concentration was applied, further in vivo experiment will be needed to prove it's safety.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.4
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• pp.349-359
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• 2010

Objective: This study was performed to compare the clinical outcome of elective single embryo transfer (eSET) performed at the cleavage stage to that of elective double embryo transfer (eDET). Methods: Of the women less than 36 years old who visited Daegu Maria from January 2008 to April 2009, the only women (n=330) with more than 8 mm of endometrial thickness and at least one good quality embryo, who were treated with GnRH agonist long protocol, were included in this study. After information about complications that can arise by multiple embryo transfer, either eSET or eDET was conducted by their request (167 and 163, respectively).Results: The implantation rate of eSET group was significantly higher than that of eDET group (53.9% vs. 40.2%, p<0.01). The twin pregnancy rate of eSET group was significantly lower than that of eDET group (1.1% vs. 32.3%, p<0.001). However, there were no significant differences between two groups in the clinical pregnancy (53.3% vs. 60.7%, p=0.172), ongoing pregnancy (47.3% vs. 54.6%, p=0.185) and live birth rates (44.9% vs. 50.9%, p=0.275). The number of the surplus embryos which developed to the blastocyst stage and cryopreserved at that stage was significantly higher in eSET group than that of eDET group ($3.2{\pm}2.6$ vs. $2.1{\pm}2.4$, p<0.001). Conclusion: These results suggest that eSET should reduce significantly the multiple baby pregnancy without decreasing the whole pregnancy rate in women with less than 36 years old.

• Tuberculosis and Respiratory Diseases
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• v.44 no.3
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• pp.547-558
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• 1997

Background : Tumor necrosis factor(TNF) has been considered as an important candidate for cancer gene therapy based on its potent anti-tumor activity. However, since the efficiency of current techniques of gene transfer is not satisfactory, the majorities of current protocols is aiming the in vitro gene transfer to cancer cells and re-introducing genetically modified cancer cells to hoot. In previous study, it was shown that TNF-sensitive cancer cells transfected with TNF-$\alpha$ cDNA would become highly resistant to TNF. Understanding the mechanisms of TNF-resistance in TNF-$\alpha$ gene transfected cancer cells would be an important step for improving the efficacy of cancer gene therapy as well as for better understandings of tumor biology. This study was designed to evaluate the role of new protective protein synthesis in the acquired resistance to TNF of TNF-$\alpha$ gene transfected cancer cells. Method : We transfected TNF-$\alpha$ c-DNA to WEHI164, a murine fibrosarcoma cell line, using retroviral vector(pLT12SN(TNF)) and confirm the expression of TNF with PCR, ELISA, MIT assay. Then we determined the TNF resistance of TNF gene transfected cells(WEHI164-TNF) and the changes of TNF sensitivities after treatments with actinomycin D(transcription inhibitor) and cycloheximide ( translation inhibitor). Results : WEHI164 which was sensitive to TNF became resistant to TNF after being transfected with TNF-$\alpha$ gene and the resistance to TNF was partially reversed after treatment with actinomycin D, but not with cycloheximide. Conclusion : The acquired resistance to TNF after TNF-$\alpha$ gene transfection may be associated with synthesis of some protective proteins.