The purpose of this study was to evaluate for the cytotoxicity of root canal sealers - Tubliseal, N2, AH26, and Sealapex - on fibroblasts cultivated from human periodontal ligament. Succinate dehydrogenase activity test and $^{51}Cr$ release test were performed to evaluate the shortterm cytotoxicity. According to l,3 and 6 days vital cell count and $^{14}C$-leucine incorporation rate to fibroblasts were evaluated. The results of this study were as follows: 1. In the test of SDH activity by millipore filter method, Sealapex showed mild cytotoxicity but Tubliseal, N2 and AH26 showed severe cytotoxicity. 2. In $^{51}Cr$ release test, Tubliseal was the most cytotoxic sealer tested, and rank ordered the relative cytotoxicity of the other sealers tested as follows: AH26, N2, Sealapex. 3. In the test of viable cell count, cytotoxicity in Tubliseal was continued because vital cell number reduced with time. Because vital cell was not showed in N2 and AH26 at 1 day, it was recognized that N2 and AH26 exhibited severe cytotoxicity. In Sealapex, vital cell number increased remarkably with time, so it showed that cytotoxicity decreased with time. 4. In $^{14}C$-leucine incorporation rate test, protein sythesis was not produced in Tubliseal, N2 and AH26 after 3 days, it showed that cytotoxicity in Tubliseal, N2 and AH26 was severe. Although protein synthesis in Sealapex decreased with time, it continued after 6 days. Therefore Sealapex has been exhibited mild cytotoxicity.
Galvez, Laura;Gil-Serna, Jessica;Garcia, Marta;Iglesias, Concepcion;Palmero, Daniel
The Plant Pathology Journal
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v.32
no.5
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pp.388-395
/
2016
The most serious aerial disease of garlic is leaf blight caused by Stemphylium spp. Geographical variation in the causal agent of this disease is indicated. Stemphylium vesicarium has been reported in Spain, whereas S. solani is the most prevalent species recorded in China. In this study, Stemphylium isolates were obtained from symptomatic garlic plants sampled from the main Spanish production areas. Sequence data for the ITS1-5.8S-ITS2 region enabled assignation of the isolates to the Pleospora herbarum complex and clearly distinguished the isolates from S. solani. Conidial morphology of the isolates corresponded to that of S. vesicarium and clearly discriminated them from S. alfalfae and S. herbarum on the basis of the size and septation pattern of mature conidia. Conidial morphology as well as conidial length, width and length:width ratio also allowed the Spanish isolates to be distinguished from S. botryosum and S. herbarum. Control of leaf blight of garlic is not well established. Few studies are available regarding the effectiveness of chemical treatments to reduce Stemphylium spp. incidence on garlic. The effectiveness of nine fungicides of different chemical groups to reduce Stemphylium mycelial growth in vitro was tested. Boscalid + pyraclostrobin (group name, succinate dehydrogenase inhibitors + quinone outside inhibitors), iprodione (dicar-boximide), and prochloraz (demethylation inhibitors) were highly effective at reducing mycelial growth in S. vesicarium with $EC_{50}$ values less than 5 ppm. In general, the effectiveness of the fungicide was enhanced with increasing dosage.
Lee Young Min;Jung Myeong Ho;Lee Yeon Sook;Song Jihyun
Journal of Nutrition and Health
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v.37
no.9
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pp.786-793
/
2004
Women with menopause or rats with ovariectomy is associated with increased body weight, body fat and insulin resistance, which are components of metabolic syndrome. Increased prevalence of metabolic syndrome after menopause might be associated with mitochondrial dysfunction, since mitochondrial oxidative and phosphorylation activity is strongly correlated with insulin sensitivity. Although estradiol replacement prevents the metabolic syndrome, harmful effect of estradiol hampers the casual usage to prevent the metabolic syndrome. It has been reported that genistein has a mild estrogenic activity, decreases fat mass in mice and has an antidiabetic role in diabetic rats. Although insulin resistance is closely related to mitochondrial functions, there has not been yet any study in regard to the effect of dietary genistein on mitochondrial function in the insulin resistant female subjects induced by ovariectomy or similar situation. The present study investigated whether the supplementation of genistein in the high fat diet affected the mitochondrial function of high fat fed ovariectomized rats. Female Sprague Dawley rats (8 weeks old) were assigned to the following groups: sham-operated+ high fat diet (S, n=6); sham-operated + high fat diet with 0.1% genistein (S + G, n=7); ovariectomized + high fat diet (OVX, n=8); ovariectomized + high fat diet with 0.1% genistein (OVX+ G, n=8). Ovariectomy significantly increased body weight compared with S group. Genistein consumption in ovariectomized (OVX + G) rats decreased body weight gain compared with OVX rats. Liver weights were increased by ovariectomy. The hepatic mitochondrial protein density expressed as mg per g liver was lower in the OVX group than in the S group. However, OVX + G group showed the increased mitochondrial protein density similar to the level of S group. When mRNA levels of genes related to mitochondria such as peroxisome proliferator-activated receptor ${\gamma}$ coactivator 1 (PGC-1) and cytochrome c oxidase subunit III (COX III) were measured, there were decreases in the mRNA levels of PGC-1 and COX III in S + G, OVX and OVX + G group. The activity of cytochrome c oxidase was not different between groups. We could observe the decrease in succinate dehydrogenase (SDH) activity per g liver in OVX rats. Genistein supplement increased SDH activity. In conclusion, genistein supplementation to the OVX rats enhanced mitochondrial function by increasing mitochondrial protein density and SDH activity. The improvement in mitochondrial function by genistein can contribute to the improvement in metabolic syndrome.
Jeong, Jae Hoon;Kim, Jeeyoung;Kim, Jeongwoon;Heo, Hye-Ryeon;Jeong, Jin Seon;Ryu, Young-Joon;Hong, Yoonki;Han, Seon-Sook;Hong, Seok-Ho;Lee, Seung-Joon;Kim, Woo Jin
Tuberculosis and Respiratory Diseases
/
v.80
no.3
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pp.247-254
/
2017
Background: Airway epithelial cells are the first line of defense, against pathogens and environmental pollutants, in the lungs. Cellular stress by cadmium (Cd), resulting in airway inflammation, is assumed to be directly involved in tissue injury, linked to the development of lung cancer, and chronic obstructive pulmonary disease (COPD). We had earlier shown that ACN9 (chromosome 7q21), is a potential candidate gene for COPD, and identified significant interaction with smoking, based on genetic studies. However, the role of ACN9 in the inflammatory response, in the airway cells, has not yet been reported. Methods: We first checked the anatomical distribution of ACN9 in lung tissues, using mRNA in situ hybridization, and immunohistochemistry. Gene expression profiling in bronchial epithelial cells (BEAS-2B), was performed, after silencing ACN9. We further tested the roles of ACN9, in the intracellular mechanism, leading to Cd-induced production, of proinflammatory cytokines in BEAS-2B. Results: ACN9 was localized in lymphoid, and epithelial cells, of human lung tissues. ACN9 silencing, led to differential expression of 216 genes. Pathways of sensory perception to chemical stimuli, and cell surface receptor-linked signal transduction, were significantly enriched. ACN9 silencing, further increased the expression of proinflammatory cytokines, in BEAS-2B after Cd exposure. Conclusion: Our findings suggest, that ACN9 may have a role, in the inflammatory response in the airway.
With 32 fungicides, it was examined the inhibitory effects on the mycelial growth of Alternaria dauci KACC42997 causing Alternaria leaf blight of carrot. Showing the results of the agar dilution method, the fungicides belonging to C2, C5, G1, E2, and E3 group were excellent in inhibiting mycelial growth. Protective fungicides belonging to M group, except for iminoctadine tris-albesilate, and pyraclostrobin belonging to C3 group were effective in inhibiting spore germination of pathogens. The fungicides included into C2 group inhibiting succinate dehydrogenase activity and the G1 group inhibiting demethylase activity showed the excellent inhibitory effect on mycelial growth but the inhibitory effect of spore germination was very low. However, fluazinam belonging to C5 group was excellent in inhibiting spore germination as well as mycelial growth. Especially, when 100 ㎍/ml of fluxapyroxad belonging to the C2 group was treated, 47.1% of spore formation was inhibited on the medium. In comparison of the resistance factors of 3 fungicide groups, as G, C, and E group, in populations of A. dauci isolates collected from Gumi, Pyeongchang, and Jeju, resistance factor in the population of Jeju was the lowest. However, two isolates resistant to fludioxonil belonging to E2 group were found in the isolate group of Pyeongchang, and both showed cross-resistance to iprodione and procymidone.
The purpose of this study was to evaluate for the cytotoxicity of glass-ionomer cement liners(GC liningcement, Ketac-bond, Vitrebond and Fuji lining LC) on the fibroblasts cultured from human pulp. The fibroblasts were cultured in DMEM-10% FBS medium. The measurement of pH, succinate dehydrogenase (SDH) activity test and $^{51}Chromium$ release test were performed. Viable cell count and $^{14}C$-leucine incorporation rate were evaluated following culture time of 2, 4 and 6 days. The results of this study were as follows : 1. The pH in all cements was to be neutralized as time elapsed, and Fuji lining LC was the lowest pH value among them. 2. SDH activity was more inhibited in GC lining cement and Vitrebond than Ketac-bond and Fuji lining LC with the setting process, and GC lining cement and Ketac-bond were reduced after 5 minute's setting and then elevated as time elapsed. 3. In SDH activity test following exposure time, the activity in Vitrebond, GC lining cement and Fuji lining LC was inhibited with increased exposure time, but it was fairly constant in Ketac-bond. 4. Overall the liquid component was more inhibited than the powder component of glass-ionomer cement in SDH activity test. 5. In $^{51}Cr$-release test, Fuji lining LC was the most released of all the cements tested and followed by : Vitrebond, Ketac-bond, GC lining cement. 6. In viable cell count, the number of cells increased as the culture day proceeded in Ketac-bond, but they decreased in GC lining cement. Fuji lining LC was only observed after 2 days culture and there was not observed the whole culture days in Vitrebond. 7. In $^{14}C$-leucine incorporation rate test, protein synthesis was decreased with the number of culture days in GC lining cement, Vitrebond and Fuji lining LC, but it was followed that of control in Ketacbond.
Fluxapyroxad is classified as carboxamide fungicide that inhibits succinate dehydrogenase in complex II of mitochondrial respiratory chain, which results in inhibition of mycelial growth within the fungus target species. This study was carried out to assure the safety of fluxapyroxad residues in agricultural products by developing an official analytical method. A new, reliable analytical method was developed and validated using High Performance liquid Chromatograph-UV/visible detector (HPLC-UVD) for the determination of fluxapyroxad residues. The fluxapyroxad residues in samples were extracted with acetonitrile, partitioned with dichloromethane, and then purified with silica solid phase extraction (SPE) cartridge. Correlation coefficient($R^2$) of fluxapyroxad standard solution was 0.9999. The method was validated using apple, pear, peanut, pepper, hulled rice, potato, and soybean spiked with fluxapyroxad at 0.05 and 0.5 mg/kg. Average recoveries were 80.6~114.0% with relative standard deviation less than 10%, and limit of detection (LOD) and limit of quantification (LOQ) were 0.01 and 0.05 mg/kg, respectively. All validation parameters were followed with Codex guideline (CAC/GL 40). LC-MS (Liquid Chromatograph-Mass Spectrometer) was also applied to confirm the analytical method. Base on these results, this method was found to be appropriate fluxapyroxad residue determination and can be used as the official method of analysis.
Kim, Sung-Kon;Kim, Nam-Kuk;Yoon, Du-Hak;Kim, Tae-Hun;Yang, Boo-Keun;Lee, Hyun-Jeong
Journal of Animal Science and Technology
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v.52
no.4
/
pp.265-270
/
2010
Adipogenesis has been one of the most intensely studied models of cellular differentiation. During adipogenesis, differential expression of many adipogenesis related genes lead to profound changes in cellular, morphological, and physiological characteristics of the differentiating cells. The aim of the present study was to examine the expression levels of adipogenic candidate genes, cAMP early repressor (ICER), nephroblastoma over-expressed protein (NOV), heat shock protein beta 1 (HSPB1) and succinate dehydrogenase (SDH), during adipogenesis of bovine mesenchymal stem cells (BMSC). The BMSC were cultured in DMEM / low glucose medium with adipogenic inducers for 6 days and the expression of various candidate genes which seemed related to adipogenesis were measured by real-time PCR. This study showed that the expression of peroxisome proliferator activated receptor ${\gamma}$(PPAR${\gamma}$) and fatty acid binding protein 4 (FABP4) genes as adipogenic indicators were increased to 3.11 and 3.11 folds on day 6 than on day 0, respectively (p<0.05). To determine whether candidate genes were related to adipogenesis, the expression levels of ICER, NOV, HSPB1, and SDH genes were measured during adipogenesis in BMSC. Our results showed that the expression level of ICER gene was significantly increased to 4.12 folds (0.01729 vs. 0.07138; p<0.05), whereas NOV, HSPB1, and SDH genes were decreased to 2.89, 3.18 and 2.36 folds, respectively, on day 6 when compared to day 0. These results suggest that these candidate genes have stimulatory or inhibitory effects on adipogenesis in BMSC, indicating that these genes may be directly or indirectly related to the adipogenic event of adipose precursor cells.
Lim, Seung-Hee;Do, Jung-Ah;Park, Shin-Min;Pak, Won-Min;Yoon, Ji Hye;Kim, Ji Young;Chang, Moon-Ik
Journal of Food Hygiene and Safety
/
v.32
no.4
/
pp.298-305
/
2017
Benzovindiflupyr is a new pyrazole carboxamide fungicide that inhibits succinate dehydrogenase of mitochondrial respiratory chain. This study was carried out to develop an analytical method for the determination of benzovindiflupyr residues in agricultural commodities using LC-MS/MS. The benzovindiflupyr residues in samples were extracted by using acetonitrile, partitioned with dichloromethane, and then purified with silica solid phase extraction (SPE) cartridge. Correlation coefficient ($r^2$) of benzovindiflupyr standard solution was 0.99 over the calibration ranges ($0.001{\sim}0.5{\mu}g/mL$). Recovery tests were conducted on 5 representative agricultural commodities (mandarin, green pepper, potato, soybean, and hulled rice) to validate the analytical method. The recoveries ranged from 79.3% to 110.0% and then relative standard deviation (RSD) was less than 9.1%. Also the limit of detection (LOD) and limit of quantification (LOQ) were 0.0005 and 0.005 mg/kg, respectively. The recoveries of interlaboratory validation ranged from 83.4% to 117.3% and the coefficient of variation (CV) was 9.0%. All results were followed with Codex guideline (CAC/GL 40) and Ministry of Food and Safety guideline (MFDS, 2016). The proposed new analytical method proved to be accurate, effective, and sensitive for benzovindiflupyr determination and would be used as an official analytical method.
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.
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