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

• Journal of Yeungnam Medical Science
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• v.23 no.1
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• pp.26-35
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• 2006

Background: Luteolin, a flavone found in various Chinese herbal medicines is known to possess anti-inflammatory properties through its ability to inhibit various proinflammatory signaling pathways including NF-${\kappa}B$ and p38 MAPK. In this study, we investigated the potential therapeutic effect of luteolin on dextran sodium sulfate (DSS)-induced colitis. Materials and Methods: We used a transgenic mouse model expressing the enhanced green fluorescent protein (EGFP) under the transcriptional control of NF-${\kappa}B$ $cis$-elements. C57BL/6 NF-${\kappa}B^{EGFP}$ mice received 2.5% DSS in their drinking water for six days in combination with daily luteolin administration (1mg/kg body weight, 0.1ml vol, intragastric) or vehicle. NF-${\kappa}B$ activity was assessed macroscopically with a Charge-Coupled Device (CCD) camera and microscopically by confocal analysis. Results: A significant increase in the Disease Activity Index (DAI), histological score (p<0.05), IL-12 p40 secretion in colonic stripe culture (p<0.05) and EGFP expression was observed in luteolin and/or DSS-treated mice compared to water-treated mice. Interestingly, a trend toward a worse colitis (DAI, IL-12p40) was observed in luteolin-treated mice compared to non-treated DSS-exposed mice. In addition, EGFP expression (NF-${\kappa}B$ activity) strongly increased in the luteolin-treated mice compared to control mice. Confocal microscopy showed that EGFP positive cells were primarily lamina propria immune cells. Conclusions: These results suggest that luteolin is not a therapeutic alternative for intestinal inflammatory disorders derived for primary defects in barrier function. Thus, therapeutic intervention targeting these signaling pathways should be viewed with caution.

• Journal of Animal Science and Technology
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• v.50 no.6
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• pp.807-818
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• 2008

This study was conducted to prove the optimum feeding method of Lactobacillus in broiler chickens by investigating the intestinal viability of ingested Lactobacillus and the effect of feeding levels and frequency of Lactobacillus on growth performance in broiler chickens. In experiment 1, A total of one hundred, 5 weeks old male broiler chickens(Abor Acre) were fed Lactobacillus reuteri avibro2 expressed green fluorescent protein(GFP) at 104cfu/g diet to investigate the retention time of ingested Lactobacillus in the intestine for 1 day. The percentage of Lactobacillus expressed GFP in intestinal contents was 26% at 1 day after fed Lactobacillus expressed GFP. The percentage of Lactobacillus expressed GFP in intestinal contents was decreased in length of time. In experiment 2, A total of four hundred eighty, 1-d-old male broiler chicks(Abor Acre) were randomly divided into 4 groups with 4 replicates of 30 birds each to prove the optimum feeding level of Lactobacillus. The treatments were control(free antibiotics), Lactobacillus reuteri avibro2 5.0×10cfu/mL, 5.0×103cfu/mL, and 5.0×105cfu/mL. The final body weight and body wight gain of Lactobacillus reuteri avibro2 5.0×103cfu/mL were the highest in all groups(P<0.05). Feed conversion ratio was not significantly difference among the groups. The number of intestinal lactic acid bacteria in Lactobacillus treated groups tended to be improved or significantly increased as compared to that of control(P<0.05). Protein and fat digestibility in Lactobacillus 5.0×103cfu/mL and 5.0×105cfu/mL treated groups were significantly improved(P<0.05). No significant differences were observed on the availability of dry matter and crude ash in Lactobacillus treatments compared to those of control. In experiment 3, A total of six hundred 1-d-old male broiler chicks(Abor Acre) were randomly divided into 4 groups with 4 replicates of 30 birds each and were fed Lactobacillus reuteri avibro2 at intervals of 1, 2, 3, and 5 day for five weeks. Feeding level of Lactobacillus was 5.0×103cfu/mL The final body weight and body wight gain of Lactobacillus reuteri avibro2 5.0×103cfu/mL were the highest in all groups(P<0.05). The final body weight and body weight gain were significantly increased, when Lactobacillus was fed at intervals of 1 days, or 2 days. There were no significant differences in feed intake and feed conversion ratio among the all groups. The number of intestinal lactic acid bacteria in Lactobacillus treated groups tended to be improved or significantly increased as compared to that of control(P<0.05). No significant differences were observed on the number of coliform bacteria and Salmonella of ileum and cecum. Consequently, supplemental Lactobacillus influenced positive effects on the growth performance, nutrient availability and intestinal microflora. The optimum feeding level of Lactobacillus was 5.0×103cfu/mL, and the constant feeding of Lactobacillus was effective.