• Journal of the Korean Society of Food Science and Nutrition
• /
• v.41 no.4
• /
• pp.437-442
• /
• 2012

Red yeast rice (RER) has been used in China for centuries for its medicinal properties and is an increasingly popular alternative lipid-lowering treatment. This study was carried out to estimate the antioxidant properties of RER extracts. The ethyl acetate extract exhibited the DPPH radical scavenging activity of 85% at 0.2 mg/mL and $IC_{50}$ 0.13 mg/mL. A significant proportion of hydroxyl radicals in a cuvette were scavenged: 44.2% at 2.5 ${\mu}g$/mL, 74.1% at 5.0 ${\mu}g$/mL, and >100% at 10 ${\mu}g$/mL. The $HepG_2$ cells pre-treated with RER ethyl acetate extract reduced the hydroxyl radicals significantly compared to the control cells. Oxidative DNA damage was measured using a Comet assay. The RER ethyl acetate extract did not induce any DNA damage per se, and appeared to enhance the resistance to DNA damage caused by an oxidant challenge with $H_2O_2$, whereas lovastatin increased the level of DNA damage in the cells in both the unstressed (no oxidant) and those stressed with $H_2O_2$. The relative gene expression of the antioxidant enzymes in $HepG_2$ cells were also affected by the RER ethyl acetate extract. The $HepG_2$ cells were pre-incubated with the RER ethyl acetate extract, and then stressed with $H_2O_2$ or left unstressed (no oxidant). In the unstressed cells, superoxide dismutase (Cu/Zn SOD) and glutathione peroxidase (GPx) were increased significantly 3.25-fold and 2.67-fold, respectively, whereas in the stressed cells, the catalase (CAT) level was increased by 4.64-fold and 7.0-fold at 5 ${\mu}g$/mL and 10 ${\mu}g$/mL, respectively, compared to those of the control. From these results, RER appears to be effective in suppressing oxidative stress.

• Maxillofacial Plastic and Reconstructive Surgery
• /
• v.28 no.5
• /
• pp.375-395
• /
• 2006

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 the korean academy of Pediatric Dentistry
• /
• v.29 no.3
• /
• pp.389-396
• /
• 2002

The purpose of this study was to compare the microtensile bonding strength of chemomechanically excavated dentin($Carisolv^{TM}$) to conventional caries removal(bur). The following adhesive systems were used; AB: All-Bond 2(3M, USA), PB: Prime & Bond 2.1(Dentsply, DE), AQ: AQ Bond(sun medical, Japan). 42 human molars with occlusal caries were assigned to 6 groups. Sequential caries removal was controlled with laser fluorescence. Each group was devided as follows; group A, B, C were $Carisolv^{TM}$ applied, group D,E,F were bur used. In group A and D, AB was used as a dentin adhesive. group B,E and group C,F was AQ and AQ was used each. The cavity was filled with composite resin(Z-100). The specimens were sectioned vertically into multiple serial 0.7 mm thick slabs. And then those slabs were sectioned into rectangular parts under 0.7 mm width. Finally 0.7-1.0 mm a right hexahedron shape stick become. Microtensile bonding test was carried out with testing apparatus at cross-head speed of $0.5\;mm/min^{-1}$ and fractured surfaces were observed with scanning electron microscope(JSM-6400, Jeol, Japan). The obtained results were summarized as follows ; 1. In the group of caries removal with $Carisolv^{TM}$, micro-tensile bonding strength decreased to $75.8{\sim}80$ percent of bur used group. 2. In the group of caries removal with $Carisolv^{TM}$, decreased degree of micro-tensile bonding strength is not so different in 3 kinds of dentin adhesives(p<0.05). 3. In the group of caries removal with $Carisolv^{TM}$, microtensile bonding strength of AB, PB, AQ was 32.6MPa(2.4), 30.1Mpa (1.8), 21.2Mpa(1.9). 4. In the group of caries removal with Bur and $Carisolv^{TM}$, microtensile bonding strength of AQ was significantly lower than that of AB and PB(p<0.01).

• Journal of Bio-Environment Control
• /
• v.27 no.4
• /
• pp.285-293
• /
• 2018

This study aimed to evaluate the effect of height suppression of cucumber and tomato plug seedlings as affected by mechanical stimulus using brushing as environment-friendly method. Cucumber (Cucumis sativus L. 'Joeunbaekdadagi') and tomato (Solanum lycopersicum L. 'Mini Chal') seeds were sown in 40-cell plug trays ($54{\times}27.5{\times}5cm$) filled with growing medium on Oct. 9, 2017. The cultivation environment in a venlo-type glasshouse was maintained as cultivation temperature range of $15-25^{\circ}C$ and the relative humidity of $50{\pm}10%$. Nontreatment and diniconazole ($7.5mg{\cdot}L^{-1}$) application at 15 days after sowing were used as the control. In addition, brushing treatments in cucumber and tomato were applied interval of 2, 4 or 6 hrs for 15 and 20 days, respectively. Plant height, hypocotyl length, and internode length were inhibited for cucumber and tomato in the diniconazole treatment than in the control. The leaf size was reduced, both cucumber and tomato, while the SPAD increased under the diniconazole treatment. However, stem diameter of cucumber was the thickest in the 2 hrs brushing interval treatment. Fresh weights of shoot and root were the significantly lowest in the diniconazole treatment. Application of brushing improved seedlings quality by promoting dry weights of shoot and root, and compactness of tomato seedlings. The chlorophyll fluorescence of tomato seedlings drastically decreased with 2 hrs treatment, indicating that mechanical stress by brushing treatment. The relative growth rate of tomato seedlings was significantly lower in the diniconazole treatment, but cucumber seedlings were not significantly different in all treatments. As a results, height suppression of cucumber and tomato seedlings was best achievement in the diniconazole treatment by the chemical as growth regulator. In an environment-friendly point of view, however, it is considered that 2 hrs brushing interval treatment can be the applicability for replacing the chemical methods in plug seedling growth of cucumber and tomato.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.36 no.3
• /
• pp.186-196
• /
• 2010

Introduction: The first aim of this study was to isolate the dental tissue-derived stem cells from the dental follicle (DF), dental pulp (DP), and root apical papilla (RAP) of the extracted wisdom teeth. Second was to evaluate their characterization with the expressions of transcription factors and cell surface markers. Finally, their ability of the in vitro multi-lineage differentiations into osteogenic and adipogenic cells were compared, respectively. Materials and Methods: Dental tissues, including dental follicle, dental pulp, and root apical papilla, were separated in the extracted wisdom teeth. These three dental tissues were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with supplements, respectively. After passage 3, the homogeneous shaped dental tissue-derived cells were analyzed the expression of transcription factors (Oct-4, Nanog and Sox-2) and cell surface markers (CD44, CD90 and CD105) with reverse transcription polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. In order to evaluate in vitro multi-lineage differentiations, the culture media were changed to the osteogenic and adipogenic induction mediums when the dental tissue-derived cells reached to passage 3. The characteristics of these three dental tissue-derived cells were compared with immunohistochemistry. Results: During primary culture, heterogenous and colony formatted dental tissue-derived cells were observed in the culture plates. After passage 2 or 3, homogenous spindle-like cells were observed in all culture plates. Transcription factors and mesenchymal stem cell markers were positively observed in all three types of dental tissue-derived cells. However, the quantity of expressed transcription factors was most large in RAP-derived cells. In all three types of dental tissue-derived cells, osteogenic and adipogenic differentiations were observed after treatment of specific induction media. In vitro adipogenic differentiation was similar among these three types of cells. In vitro osteogenic differentiation was most strongly and frequently observed in the RAP-derived cells, whereas rarely osteogenic differentiation was observed in the DP-derived cells. Conclusion: These findings suggest that three types of human dental tissue-derived cells from extracted wisdom teeth were multipotent mesenchymal stem cells, have the properties of multi-lineage differentiations. Especially, stem cells from root apical papilla (SCAP) have much advantage in osteogenic differentiation, whereas dental follicle cells (DFCs) have a characteristic of easy adipogenic differentiation.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.28 no.4
• /
• pp.613-619
• /
• 2001

The aims of this study were to compare the accuracy, sensitivity and specificity of cnventional visual examination, radiography and a new laser fluorescence method, KaVo Diagnodent, for the detection of occlusal caries lesions. One hundred sound human premolars and molars which had no restorations or interproximal cavities were tested by three methods. Tooth lesions depth was assessed at histologic examination using Caries detector dye The following results were obtained. 1. Diagnodent show 7.8 in sound tooth, 25.4 in initial caries, 30.5 in enamel caries, and 53.8 in dentin caries with average score 2. Spearman and Pearson relation coefficient was high between tooth-specimen test with dye and Diagnodent(0.736, 0.619), visual examination(0.664, 0.666), and was low between tooth-specimen test with dye and radiographic examination(P<0.01, total) 3. Accuracy of occlusal caries was highest on Diagnodent(65%) and lowest on radiographic examination(35%) 4. In initial caries, the sensitivity and specificity of Diagnodent method was the highest. In enamel caries, the sensitivity of visual examination was the highest and specificity of Diagnodent method was the highest. In dentinal caries, the sensitivity and specificity of Diagnodent method was the highest and sensitivity of visual examination was the lowest.

• Clinical and Experimental Reproductive Medicine
• /
• v.37 no.3
• /
• pp.217-229
• /
• 2010

Objective: Apoptosis plays an important role for the maintenance of the normal pregnancy. Expression of X-linked inhibitor of apoptosis (XIAP) is able to effectively prevent apoptosis and controls trophoblast cells death throughout placental development, but it is still unknown in the function of XIAP in trophoblast cells exposed to hypoxic condition, which is one of the factors causing preeclampsia. Therefore, we conducted to compare XIAP expression in normal and pre-eclamptic placenta tissues and analyzed the function of XIAP in HTR-8/SVneo trophoblast cell line exposed to hypoxic condition. Methods: The expression of XIAP was analyzed in placental tissues from the following groups of patients (none underwent labor): 1) term normal placenta (n=15); 2) term with pre-eclamptic placeneta (n=15); and 3) pre-term with pre-eclamptic placenta (n=11) using semi-quantitative RT-PCR, immunohistochemistry, and Western blot. In order to evaluate the function of XIAP in HTR-8/SVneo trophoblast cells under hypoxic condition, HIF-$1{\alpha}$ plasmids, and hypoxic condtion were transfected and treated into HTR-8/SVneo trophoblast cells for 24 hours, respectively. Results: We observed that XIAP are expressed in the syncytiotrophoblasts and syncytial knot of placental villi. The expression of XIAP was significantly decreased in preeclamptic placenta tissues than in normal placenta tissues without labor (p<0.05). Furthermore, we confirmed the XIAP expression in HTR-8/SVneo trophbolast cells exposed to hypoxia was translocated from cytoplasm into nucleus and decreased XIAP by hypoxic condition induced apoptosis in HTR-8/SVneo trophoblast cells through up-regulation of pro-apoptotic proteins. Conclusion: These results suggest that the expression of XIAP is involved in placental development as well as decreased expression of XIAP by hypoxia is associated with pre-eclampsia through inducing trophoblast cells apoptosis.

• Journal of Life Science
• /
• v.16 no.4
• /
• pp.571-578
• /
• 2006

HtrA2/Omi, a mitochondrial trypsin-like serine protease, is pivotal in regulating apoptotic cell death. Several lines of recent evidence suggest that HtrA2 is associated with the pathogenesis of neurodegenerative disorders; however, the physiological function of HtrA2 still remains elusive. For studying physiological function of HtrA2 in depth, it is necessary to develop a suitable expression system in the model animal. We therefore utilized the zebrafish as a model animal to establish expression of human HtrA2 (hHtrA2) in vivo. For expression of mature HtrA2 as GFP fusion in zebrafish embryos, the HtrA2 (WT) or (S306A) cDNAs with the C-terminal GFP tag were inserted into the pCS2+ plasmid. Expression patterns of HtrA2 in HEK293 cells were first monitored by immunofluorescence staining and immunoblot assays, showing approximately 64 kDa of the HtrA2-GFP fusion proteins. Subsequently, the hHtrA2 plasmid DNA or in vitro transcribed mRNA was microinjected into zebrafish embryos. The expression patterns of HtrA2 in Zebrafish embryos were monitored by GFP fluorescence in 24 hours-post-fertilization (hpf). Although expression patterns of HtrA2-GFP in developing embryos were different between the injected DNA and mRNA, both nucleic acids revealed good expression levels to further study the physiological role of HtrA2 in vivo. This study provides a suitable condition for expressing hHtrA2 in the zebrafish embryos as well as a method for generating useful system to investigate physiological properties of the specific human genes.

• Journal of Life Science
• /
• v.27 no.12
• /
• pp.1445-1451
• /
• 2017

Mitochondria play a central role in energy generation by using electron transport coupled with oxidative phosphorylation. They also participate in other important cellular functions including metabolism, apoptosis, signaling, and reactive oxygen species production. Therefore, mitochondrial dysfunction is known to contribute to a variety of human diseases. Furthermore, there are various inherited diseases of energy metabolism due to mitochondrial DNA (mtDNA) mutations. Unfortunately, therapeutic options for these inherited mtDNA diseases are extremely limited. In this regard, mitochondrial replacement techniques are taking on increased importance in developing a clinical approach to inherited mtDNA diseases. In this study, green fluorescence protein (GFP)-tagged mitochondria were isolated by differential centrifugation from a mammalian cell line. Using microinjection technique, the isolated GFP-tagged mitochondria were then transferred to bovine oocytes that were triggered for early development. During the early developmental period from bovine oocytes to blastocysts, the transferred mitochondria were observed using fluorescent microscopy. The microinjected mitochondria were dispersed rapidly into the cytoplasm of oocytes and were passed down to subsequent cells of 2-cell, 4-cell, 8-cell, morula, and blastocyst stages. Together, these results demonstrate a successful in vitro transfer of isolated mitochondria to oocytes and provide a model for mitochondrial replacement implicated in inherited mtDNA diseases and animal cloning.

• Tuberculosis and Respiratory Diseases
• /
• v.56 no.3
• /
• pp.261-268
• /
• 2004

Background : The resurgence of tuberculosis and outbreaks of multidrug resistant (MDR) tuberculosis have increased the emphasis for the development of new susceptibility testing of the Mycobacterium tuberculosis for the effective treatment and control of the disease. Conventional drug susceptibility testings, such as those using egg-based or agar-based media have some limits, such as the time required and difficulties in determining critical inhibitory concentrations, but these are still being used in many diagnostic laboratories because of no better lternatives, considering cost and accuracy. To overcome these limits, a rapid and simple method for new susceptibility testing, using live and dead assays, was applied for a bacterial cell viability assay to distinguish dead from live bacterial cells based on two-color fluorescence. Materials and Methods Strains : Forty strains were used in this study, 20 susceptible to all antituberculosis drugs and the other 20 resistant to the four first line antituberculosis drugs isoniazid, rifampicin, streptomycin and ethambutol. Antibiotics : The four antibiotics were dissolved in 7H9 broth to make the following solutions: $0.1{\mu}g\;isoniazid(INH)/m{\ell}$, $0.4{\mu}g\;rifampicin(RMP)/m{\ell}$, $4.0{\mu}g\;streptomycin(SM)/m{\ell}$ and $4.0{\mu}g\;ethambutol(EMB)/m{\ell}$. Results : Live and dead Mycobacterium tuberculosis cells fluoresced green and red with the acridin (Syto 9) and propidium treatments, respectively. These results are very well accorded with conventional drug susceptibility testing by proportional method on Lowensen-Jensen media (L-J) containing 4 drugs (INH, RMP, EMB and SM), showing a 93.7 % accordance rate in susceptible strains and 95% in resistant strains. Conclusion : The results of the drug susceptibility testing using the live and dead bacterial cell assay showed high accordance rates compared with the conventional proportion method on L-J. This finding suggests that the live and dead bacterial cell assay can be used as an alternative to conventional drug susceptibility testing for M. tuberculosis strains.