Mitochondrial dysfunction influences apoptosis and autophagy in porcine parthenotes developing in vitro     

Xu YN  Cui XS  Sun SC  Lee SE  Li YH  Kwon JS  Lee SH  Hwang KC  Kim NH 《The Journal of reproduction and development》2011,57(1):143-150

Mitochondria are important regulators of both apoptosis and autophagy. One of the triggers for mitochondrial-mediated apoptosis is the production of reactive oxygen species (ROS), which include hydrogen peroxide, superoxide, hydroxyl radical, nitric oxide and peroxynitrite. Recently, several studies have indicated that ROS may also be involved in the induction of autophagy. In the present study, we used H(2)O(2) to induce mitochondrial stress, examined apoptotic- and autophagic-related gene expression and observed LC3 protein (autophagosome presence marker) expression in porcine parthenotes developing in vitro. In porcine four-cell parthenotes cultured for 5 days in NCSU37 medium containing 0.4% BSA, the developmental rate and mitochondrial distribution did not differ from that of the group supplemented with 100 μM H(2)O(2) but was significantly decreased in the group supplemented with 500 μM H(2)O(2) (P<0.05). Transmission electron microscopy (TEM) indicated that whereas normal shaped mitochondria were observed in blastocysts from the control group, abnormal mitochondria (mitophagy) and autophagic vacuoles were observed in blastocysts from the group that received 500 μM H(2)O(2). Furthermore, addition of H(2)O(2) (100 μM and 500 μM) decreased cell numbers (P<0.05) and increased both apoptosis (P<0.05) and LC3 protein expression in the blastocysts. Real-time RT-PCR showed that H(2)O(2) significantly decreased mRNA expression of anti-apoptotic gene Bcl-xL but increased pro-apoptotic genes, Caspase 3 (Casp3) and Bak, and autophagy-related genes, microtubule-associated protein 1 light chain 3 (Map1lc3b) and lysosomal-associated membrane protein 2 (Lamp2). However, the addition of H(2)O(2) had no effect on mRNA expression levels in nuclear DNA-encoded mitochondrial-related genes, cytochrome oxidase (Cox) 5a, Cox5b and Cox6b1, in blastocysts. These results suggest that H(2)O(2) leads to mitochondrial dysfunction that results in apoptosis and autophagy, which is possibly related to porcine early embryo development.  相似文献   

Effects of activation methods on DNA synthesis and development of parthenogenetic porcine embryos   总被引：1，自引：0，他引：1  

Ock SA  Kwack DO  Mohana Kumar B  Han J  Kim SW  Rho GJ 《Reproduction in domestic animals》2011,46(6):1082-1089

This study investigated the timing of DNA synthesis and patterns of pronuclear (PN) formation during the first cell cycle, and its influence on developmental competence, velocity and proliferation index of porcine parthenote blastocysts produced by different activation treatments. Oocytes were activated as follows: electrical stimulation (EST), EST combined with 7.5 μg/ml cytochalasin B (EST + CCB), 10 μg/ml cycloheximide (EST + CHX) and 1.9 mm 6-dimethylaminopurine (EST + 6-DMAP) for 3 h. DNA synthesis and PN formation were evaluated using 1 mm 5'bromo-2'deoxy-uridne (BrdU) at 2 h intervals from 1 to 13 h or 5 to 13 h of post-activation (hpa), respectively. In EST, DNA synthesis started at 3 hpa, reached the peak at 11 hpa and decreased at 13 hpa. Treatment with 6-DMAP resulted in an early increase of DNA synthesis at 3 hpa, whereas CCB delayed DNA synthesis for 2 h. In EST and EST + 6-DMAP, most of the eggs showed 1PN, whereas, incidence of 2PN in EST + CCB was higher than 1PN. EST + CHX was observed with 1PN, 2PN and multiple PN. Blastocyst rate in EST + CCB and EST + 6-DMAP were significantly (p<0.05) higher than EST + CHX. But, the developmental velocity was not different among groups. Proliferation index of blastocysts, as indicated the number of blastomere at S-phase of the cell cycle was low in all groups. In conclusion, CCB, CHX and 6-DMAP used for producing porcine parthenogenetic embryos induced different onset of DNA synthesis and PN, but they did not affect the subsequent embryo development.  相似文献   

Low-dose fish oil consumption prevents hepatic lipid accumulation in high cholesterol diet fed mice     

Hirako S  Kim HJ  Shimizu S  Chiba H  Matsumoto A 《Journal of agricultural and food chemistry》2011,59(24):13353-13359

We examined the effects of low-dose fish oil ingestion on hepatic lipid accumulation caused after high cholesterol feeding in C57BL/6J mice. The mice were fed purified experimental diets consisting of 20 energy % (en%) safflower oil (SO or SO/CH), 2 en% fish oil + 18 en% safflower oil (2FO or 2FO/CH), or 5 en% fish oil + 15 en% safflower oil (5FO or 5FO/CH) with or without 2 weight % (wt %) cholesterol for 8 weeks. Hepatic triglyceride and total cholesterol contents were significantly lower in groups that were fed diets containing fish oil and cholesterol than in those that were fed safflower oil and cholesterol. The hepatic mRNA levels of fatty acid synthase (FAS) were lower in groups fed cholesterol or fish oil. Fatty acid oxidation-related hepatic gene expressions were higher in fish oil-fed groups. Fecal cholesterol excretion was higher in all cholesterol-fed groups; cholesterol excretion was high in groups fed fish oil and cholesterol. These results suggest that low-dose fish oil diets improve lipid metabolism by modifying the expression of lipid metabolism-related genes in the liver and increasing fecal cholesterol excretion.  相似文献   

Metabolic dependence of green tea on plucking positions revisited: a metabolomic study     

Lee JE  Lee BJ  Hwang JA  Ko KS  Chung JO  Kim EH  Lee SJ  Hong YS 《Journal of agricultural and food chemistry》2011,59(19):10579-10585

The dependence of global green tea metabolome on plucking positions was investigated through (1)H nuclear magnetic resonance (NMR) analysis coupled with multivariate statistical data set. Pattern recognition methods, such as principal component analysis (PCA) and orthogonal projection on latent structure-discriminant analysis (OPLS-DA), were employed for a finding metabolic discrimination among fresh green tea leaves plucked at different positions from young to old leaves. In addition to clear metabolic discrimination among green tea leaves, elevations in theanine, caffeine, and gallic acid levels but reductions in catechins, such as epicatechin (EC), epigallocatechin (EGC), epicatechin-3-gallate (ECG), and epigallocatechin-3-gallate (EGCG), glucose, and sucrose levels were observed, as the green tea plant grows up. On the other hand, the younger the green tea leaf is, the more theanine, caffeine, and gallic acid but the lesser catechins accumlated in the green tea leaf, revealing a reverse assocation between theanine and catechins levels due to incorporaton of theanine into catechins with growing up green tea plant. Moreover, as compared to the tea leaf, the observation of marked high levels of theanine and low levels of catechins in green tea stems exhibited a distinct tea plant metabolism between the tea leaf and the stem. This metabolomic approach highlights taking insight to global metabolic dependence of green tea leaf on plucking position, thereby providing distinct information on green tea production with specific tea quality.  相似文献   

Influenza virus neuraminidase inhibitory activity of phlorotannins from the edible brown alga Ecklonia cava     

Ryu YB  Jeong HJ  Yoon SY  Park JY  Kim YM  Park SJ  Rho MC  Kim SJ  Lee WS 《Journal of agricultural and food chemistry》2011,59(12):6467-6473

Influenza A virus infections continue to pose a major threat to humans and several animal species. Neuraminidase (NA) is one of the most promising targets for the development of drugs against influenza viruses because of its critical role in the viral life cycle. During the course of a search for NA inhibitors from edible natural sources, we found that the ethyl acetate layer of ethanol extracts of Ecklonia cava showed extremely high NA-inhibitory activity (72.1% inhibition at 30 μg/mL). Bioactivity-guided fractionation of the ethyl acetate layer yielded five phlorotannins, identified as phloroglucinol (1), eckol (2), 7-phloroeckol (3), phlorofucofuroeckol (4), and dieckol (5). The inhibitory activities of these compounds (1-5) against NAs from group-1 (A/Bervig_Mission/1/18 [H1N1], A/PR/8/34 [H1N1]) and group-2 (A/Hong Kong/8/68 [H3N2], A/Chicken/Korea/MS96/96 [H9N2]) influenza A were evaluated to determine potencies and kinetic behavior. Analyses using various in vitro influenza A virus NA assays showed that all five phlorotannin derivatives were selective NA inhibitors. Of the phlorotannins, phlorofucofuroeckol (4) exhibited the most potent inhibitory activities toward group-1 NAs (IC?? values, 4.5 and 14.7 μM), whereas dieckol (5) potently inhibited group-2 NAs. Kinetic analyses indicated that compounds 1-5 were all noncompetitive. Notably, these noncompetitive inhibitors synergized with oseltamivir to enhance the NA-inhibitory effects of oseltamivir.  相似文献   

Evaluation of the effects of thiopental, propofol, and etomidate on glomerular filtration rate measured by the use of dynamic computed tomography in dogs     

Chang J  Kim S  Jung J  Lee H  Chang D  Lee Y  Lee I  Yoon J  Choi M 《American journal of veterinary research》2011,72(1):146-151

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Effect of the use of carprofen in dogs undergoing intense rehabilitation after lateral fabellar suture stabilization     

Gordon-Evans WJ  Dunning D  Johnson AL  Knap KE 《Journal of the American Veterinary Medical Association》2011,239(1):75-80

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Efficacy of bacteriophage therapy on horizontal transmission of Salmonella gallinarum on commercial layer chickens     

Lim TH  Lee DH  Lee YN  Park JK  Youn HN  Kim MS  Lee HJ  Yang SY  Cho YW  Lee JB  Park SY  Choi IS  Song CS 《Avian diseases》2011,55(3):435-438

A Salmonella Gallinarum (SG)-specific bacteriophage isolated from sewage effluent was used to prevent horizontal transmission of SG in commercial layer chickens. Six-week-old chickens, each challenged with 5 x 10(8) colony-forming units of SG, cohabited with contact chickens treated with 10(6) plaque-forming units/kg of bacteriophage, prepared in feed additives, for 7 days before, and 21 days after challenge with SG. Mortality was observed for 3 wk after challenge and SG was periodically reisolated from the liver, spleen, and cecum of chickens. SG re-isolation from organs was decreased and a significant (P < 0.05) reduction in mortality was observed in contact chickens treated with the bacteriophage, as compared to untreated contact chickens, indicating that bacteriophage administration in feed additives significantly prevented the horizontal transmission of SG. These results provide important insights into prevention and control strategies against SG infection and suggest that the use of bacteriophages may be a novel, safe, and effectively plausible alternative to antibiotics for the prevention of SG infection in poultry.  相似文献   

Identification and virulence characterization of fowl adenoviruses in Korea     

Lim TH  Lee HJ  Lee DH  Lee YN  Park JK  Youn HN  Kim MS  Youn HS  Lee JB  Park SY  Choi IS  Song CS 《Avian diseases》2011,55(4):554-560

Since 2007, 55 adenovirus strains have been isolated from commercial chicken flocks in Korea and have been identified and the pathogenicity of these isolates was confirmed in specific-pathogen-free chickens of different age. Based on sequencing analysis of the hexon gene, 55 FAdV isolates were genetically related to the IBH-2A strain of FAdV3 (4 isolates, 99.2% to 100%), the KR5 strain of FAdV4 (22 isolates, 97.9% to 99.2%), the 764 strain of FAdV9 (11 isolates, 99.1% to 99.3%), and the 1047 strain of FAdV11 (18 isolates, > 99%). Experimental infections with four serotypes of FAdV resulted in high mortality of 18-day-old chicken embryos and 1-day-old chicks with marked liver necrosis similar to those observed in the natural outbreaks. Notably, specific hydropericardium was observed in chicks challenged with the K531 strain (serotype 4). However, 3-wk-old chickens challenged with FAdVs, regardless of serotype, did not show any clinical signs or mortality except histologic lesions of focal hepatocytic necrosis with mild lymphocytic infiltration. The results indicate that four FAdV serotypes (3, 4, 9, and 11) are the dominant serotypes of FAdVs in the Korea and are pathogenic enough to cause clinical disease in young chicks. The present investigation provides important information on the epidemiology and pathogenesis of FAdVs and highlights the importance of control strategies against FAdV infection in Korea.  相似文献   

Corn distillers dried grains with solubles in diets for growing-finishing pigs: a cooperative study     

Cromwell GL  Azain MJ  Adeola O  Baidoo SK  Carter SD  Crenshaw TD  Kim SW  Mahan DC  Miller PS  Shannon MC;North Central Coordinating Committee on Swine Nutrition 《Journal of animal science》2011,89(9):2801-2811

An experiment involving 560 crossbred pigs (28 replications of 4 to 6 pigs per pen) was conducted at 9 research stations to assess the effects of dietary concentrations of corn distillers dried grains with solubles (DDGS) on pig performance and belly firmness. Fortified corn-soybean meal diets containing 0, 15, 30, or 45% DDGS were fed in 3 phases from 33 to 121 kg of BW. A common source of DDGS containing 90.1% DM, 26.3% CP, 0.96% Lys, 0.18% Trp, 9.4% crude fat, 34.6% NDF, 0.03% Ca, and 0.86% P was used at each station. Diets were formulated to contain 0.83, 0.70, and 0.58% standardized ileal digestible (SID) Lys during the 3 phases with diets changed at 60 and 91 kg of BW, respectively. The DDGS replaced corn and soybean meal, and up to 0.172% Lys and 0.041% Trp were added to maintain constant SID concentrations of Lys and Trp in each phase. At each station, 2 pigs from each pen in 2 replications were killed and a midline backfat core was obtained for fatty acid analysis and iodine value. In most instances, there were differences among stations (P < 0.01), but the station × treatment interactions were few. Body weight gain was linearly reduced in pigs fed the greater amounts of DDGS (0 to 45%) during phase I (950, 964, 921, and 920 g/d; P < 0.01) and over the entire experimental period (944, 953, 924, and 915 g/d; P = 0.03), but ADFI (2.73, 2.76, 2.68, and 2.70 kg) and G:F (347, 347, 345, and 341 g/kg) were not affected (P = 0.15 and P = 0.33, respectively) during the entire test. Backfat depth was reduced (linear, P < 0.02) by increasing amounts of DDGS (22.5, 22.7, 21.4, and 21.6 mm), but LM area (47.4, 47.4, 46.1, and 45.4 cm(2)) was not affected (P = 0.16) by treatments. Estimated carcass fat-free lean was 51.9, 52.2, 52.4, and 52.1% for 0 to 45% DDGS, respectively (linear, P = 0.06). Flex measures obtained at 6 stations indicated less firm bellies as dietary DDGS increased (lateral flex: 11.9, 8.6, 8.4, and 6.6 cm; linear, P < 0.001; vertical flex: 26.1, 27.4, 28.2, and 28.7 cm; linear, P < 0.003). Saturated and monounsaturated fatty acid concentrations in subcutaneous fat decreased linearly (P < 0.001) and PUFA concentrations increased linearly (P < 0.001) with increasing DDGS in the diet. Iodine values in inner (61.1, 68.2, 74.7, and 82.2) and outer (67.9, 73.6, 79.6, and 85.8) backfat increased linearly (P < 0.001) as DDGS in the diet increased. In this study, feeding diets with 30 or 45% DDGS did not have major effects on growth performance, but resulted in softer bellies. Regression analysis indicated that iodine values increased 4.3 units for every 10 percentage unit inclusion of DDGS in the diet.  相似文献