Choi, Hyeong Sim;Jeong, Eun-Hui;Lee, Tae-Gul;Kim, Seo Yun;Kim, Hye-Ryoun;Kim, Cheol Hyeon
Tuberculosis and Respiratory Diseases
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v.75
no.1
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pp.9-17
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2013
Background: In cancer cells, autophagy is generally induced as a pro-survival mechanism in response to treatment-associated genotoxic and metabolic stress. Thus, concurrent autophagy inhibition can be expected to have a synergistic effect with chemotherapy on cancer cell death. Monensin, a polyether antibiotic, is known as an autophagy inhibitor, which interferes with the fusion of autophagosome and lysosome. There have been a few reports of its effect in combination with anticancer drugs. We performed this study to investigate whether erlotinib, an epidermal growth factor receptor inhibitor, or rapamycin, an mammalian target of rapamycin (mTOR) inhibitor, is effective in combination therapy with monensin in non-small cell lung cancer cells. Methods: NCI-H1299 cells were treated with rapamycin or erlotinib, with or without monensin pretreatment, and then subjected to growth inhibition assay, apoptosis analysis by flow cytometry, and cell cycle analysis on the basis of the DNA contents histogram. Finally, a Western blot analysis was done to examine the changes of proteins related to apoptosis and cell cycle control. Results: Monensin synergistically increases growth inhibition and apoptosis induced by rapamycin or erlotinib. The number of cells in the sub-$G_1$ phase increases noticeably after the combination treatment. Increase of proapoptotic proteins, including bax, cleaved caspase 3, and cleaved poly(ADP-ribose) polymerase, and decrease of anti-apoptotic proteins, bcl-2 and bcl-xL, are augmented by the combination treatment with monensin. The promoters of cell cycle progression, notch3 and skp2, decrease and p21, a cyclin-dependent kinase inhibitor, accumulates within the cell during this process. Conclusion: Our findings suggest that concurrent autophagy inhibition could have a role in lung cancer treatment.
The present study was aimed at investigating the regulatory mechanism in tissue-specific expression of insulin-like growth factor-I (IGF-I) gene. The expression of IGF-I gene was determined by a solution hybridization/RNase protection assay using total RNA prepared from rat liver or brain of various ages. The levels of IGF-I transcripts were increased in liver gradually after birth, but decreased in brain. By using an oligonucleotide (FRE) corresponding to the C/EBP binding site of the rat IGF-I exon 1, multiple forms of C/EBP${\alpha}$ and C/EBP${\beta}$ proteins, which have DNA-binding activity, were detected in the rat liver or brain. Western immunoblot and southwestern analyses show that p42$\^$C/EBP${\alpha}$/, p38$\^$C/EBP${\alpha}$/, p35$\^$C/EBP${\alpha}$/, p38$\^$C/EBP${\beta}$/, and p35$\^$C/EBP${\beta}$ form specific complexes with the IGF-I exon 1 oligonucleotide in liver nuclear extract and that p42$\^$C/EBP${\alpha}$/ and p38$\^$C/EBP${\beta}$/ form complexes in brain. These data suggest that the formation of FRE-C/EBP isoform complexes may play important roles in the tissue-specific regulation of IGF-I gene expression.
Kim, D.W.;Jang, B.G.;Kim, J.H.;Yu, D.J.;Kang, K.H.;Kang, H.G.;Na, J.C.;Kim, S.H.;Lee, D.S.;Suh, O.S.
Korean Journal of Poultry Science
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v.34
no.2
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pp.137-141
/
2007
This experiment was conducted to investigate the effects of dietary yeast (Saccharomyces exguus) supplementation on growth performance, cecal microflora and fecal ammonia gas in broiler chicks. A total of two hundred seventy, 1-d-old male broiler chicks (Ross strain) were randomly allotted to nine pens (replicates), 30 birds per pen. There were three dietary treatments with three replicates. The treatments were control (virginiamycin 0.05%+salinomycin 0.03%), Saccharomyces exguus 0.5 and 1.0%. Total body weight gain were significantly higher in Saccharomyces exguus1 1.0% treatment than the control (P<0.05). Although not significant, the yeast supplementation tended to improve the feed conversion ratio. No significant differences were observed on the numbers of cecal E. coli, Salmonella and Lactobacillus in yeast treatments compared to those of control. The production of fecal ammonia gas was significantly lower in yeast treatments than the control (P<0.05). The concentrations of fecal short chain fatty acids and volatile organic compounds were not different among the groups. These results suggest the possibility that yeast (Saccharomyces exguus) could be used as the alternative of antibiotic growth promoters by improving the performance of broiler chicks. In addition, dietary yeast could improve the environment of broiler houses by reducing fecal ammonia production.
Irawan, Agung;Hidayat, Cecep;Jayanegara, Anuraga;Ratriyanto, Adi
Animal Bioscience
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v.34
no.9
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pp.1499-1513
/
2021
Objective: The purpose of this meta-analysis was to evaluate the effect of dietary essential oils (EOs) on productive performance, nutrient digestibility, and serum metabolite profiles of broiler chickens and to compare their effectiveness as growth-promoting additives against antibiotics. Methods: Peer-reviewed articles were retrieved from Web of Science, Science Direct, PubMed, and Google scholar and selected based on pre-determined criteria. A total of 41 articles containing 55 experiments with 163 treatment units were eligible for analyses. Data were subjected to a meta-analysis based on mixed model methodology considering the doses of EOs as fixed effects and the different studies as random effects. Results: Results showed a linear increase (p<0.001) on body weight gain (BWG) where Antibiotics (FCR) and average daily feed intake decreased (p<0.001) linearly with an increasing dose of EOs. Positive effects were observed on the increased (p<0.01) digestibility of dry matter, crude protein, ether extract, and cecal Lactobacillus while Escherichia coli (E. coli) population in the cecum decreased (p<0.001) linearly. There was a quadratic effect on the weight of gizzard (p<0.01), spleen (p<0.05), bursa of fabricius (p<0.001), and liver (p<0.10) while carcass, abdominal fat, and pancreas increased (p<0.01) linearly. The dose of EOs linearly increased high density lipoprotein, glucose, protein, and globulin concentrations (p<0.01). In comparison to control and antibiotics, all type of EOs significantly reduced (p<0.001) FCR and tended to increase (p<0.1) BWG and final body weight. Cinnamaldehyde-compound was the only EOs type showing a tendency to increase (p<0.1) carcass weight, albumin, and protein of serum metabolites while this EOs together with EOs-Blend 1 decreased (p<0.01) E. coli population. Low density lipoprotein concentration decreased (p<0.05) with antibiotics and carvacrol-based compound when compared to the control group. Conclusion: This evidence confirms that EOs are suitable to be used as growth promoters and their economical benefit appears to be promising.
This study's aim is to isolate and characterize plant growth promoting Enterobacter species for the biological control of gray leaf spot in pepper. Screening was carried out from the rhizosphere of Agropyron tsukushiensi var. transiens (Hack.) Ohwi in Dok-do. Rhizobacterial isolates were partially identified by 16S rDNA sequencing and Enterobacter species were tested for plant growth promoting capabilities and the induction of systemic resistance in pepper against gray leaf spot caused by Stemphylium solani. Isolates were tested for production of indole-acetic acid and siderophore, and for phosphate solubilization. The application of isolates was effective in controlling gray leaf spot in pepper with E. asburiae (KNUC5007) and E. cancerogenes (KNUC5008 and KNUC5010) having the highest efficacy in reducing gray leaf spot severity. This is the first report of the biological control of gray leaf spot in pepper using rhizobacteria and it is hoped that this study will increase the utilization of Enterobacter species as plant growth promoters and biocontrol agents.
Pearson, Brianna;Lau, Kin H.;Allen, Alicia;Barron, James;Cool, Robert;Davis, Kelly;DeLoache, Will;Feeney, Erin;Gordon, Andrew;Igo, John;Lewis, Aaron;Muscalino, Kristi;Parra, Madeline;Penumetcha, Pallavi;Rinker, Victoria G.;Roland, Karlesha;Zhu, Xiao;Poet, Jeffrey L.;Eckdahl, Todd T.;Heyer, Laurie J.;Campbell, A. Malcolm
Interdisciplinary Bio Central
/
v.3
no.3
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pp.10.1-10.8
/
2011
Introduction: Hash functions are computer algorithms that protect information and secure transactions. In response to the NIST's "International Call for Hash Function", we developed a biological hash function using the computing capabilities of bacteria. We designed a DNA-based XOR logic gate that allows bacterial colonies arranged in a series on an agar plate to perform hash function calculations. Results and Discussion: In order to provide each colony with adequate time to process inputs and perform XOR logic, we designed and successfully demonstrated a system for time-delayed bacterial growth. Our system is based on the diffusion of ${\ss}$-lactamase, resulting in destruction of ampicillin. Our DNA-based XOR logic gate design is based on the op-position of two promoters. Our results showed that $P_{lux}$ and $P_{OmpC}$ functioned as expected individually, but $P_{lux}$ did not behave as expected in the XOR construct. Our data showed that, contrary to literature reports, the $P_{lux}$ promoter is bidirectional. In the absence of the 3OC6 inducer, the LuxR activator can bind to the $P_{lux}$ promoter and induce backwards transcription. Conclusion and Prospects: Our system of time delayed bacterial growth allows for the successive processing of a bacterial hash function, and is expected to have utility in other synthetic biology applications. While testing our DNA-based XOR logic gate, we uncovered a novel function of $P_{lux}$. In the absence of autoinducer 3OC6, LuxR binds to $P_{lux}$ and activates backwards transcription. This result advances basic research and has important implications for the widespread use of the $P_{lux}$ promoter.
Park, Seung-Moon;Mo, Ae-Young;Jang, Yong-Suk;Lee, Jae-Hwa;Yang, Moon-Sik;Kim, Dae-Hyuk
Biotechnology and Bioprocess Engineering:BBE
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v.9
no.4
/
pp.292-296
/
2004
The recombinant soluble human tumor necrosis factor-alpha (hTNF-$\alpha$) was expressed in a yeast Saccharomyces cerevisiae and its cytotoxicity was evaluated. A cDNA encoding hTNF-$\alpha$ was placed under the control of two different promoters: a glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter and a yeast hybrid ADH2-GPD promoter, consisting of alcohol dehydrogenase II (ADH2) and the GPD promoter. A Northern blot analysis revealed that, although variation in the expression level of hTNF-$\alpha$ existed among transformants, the higher expression was obtained with the GPD promoter. Expressed hTNF-$\alpha$ protein (rhTNF-$\alpha$) was successfully secreted into the culture medium, producing 2.5 mg per liter of culture filtrate, with no changes in cell growth. The bioassay for observing the cytotoxicity to the murine L929 fibroblast cell line, with serial dilution of rhTNF-$\alpha$, indicated that the secreted rhTNF-$\alpha$ was bioactive and its dose-response was improved eight to ten times over that of the E. coli-derived rhTNF-$\alpha$.
Cyclic AMP-responsive element binding protein (CREB) is known to be associated with angiogenesis. In the present study we investigated the possible role of CREB in the expression of vascular endothelial growth factor (VEGF) by mouse macrophages. Over-expression of CREB increased VEGF secretion by cells of the RAW264.7 mouse macrophage cell line. It also increased the promoter activity of a mouse reporter driven by the VEGF promoter, while a dominant negative CREB (DN-CREB) abrogated the activity, suggesting that CREB mediates VEGF transcription. Forskolin, an adenylyl cyclase activator, stimulated VEGF transcription, and the PKA inhibitor H89 abolished this effect. IFN-${\gamma}$, a potent cytokine, stimulated VEGF expression only in part through the PKA-CREB pathway. These results indicate that PKA phosphorylates CREB and so induces VEGF gene expression. An analysis of mutant promoters revealed that one of the putative CREB responsive elements (CREs), at -399 ~ -388 in the promoter, is critical for CREB-mediated VEGF promoter activity, and the significance of this CRE was confirmed by chromatin immunoprecipitation assays.
In order to study performance enhancing effects of supplementary antibiotics (avoparcin, nosiheptide and enramycin), two feeding trials were conducted. In Experiment 1, 1, 040 male Arbor Acres were reared in floor pens for 6 wk. Chicks were assigned to one of the following four treatments: Basal diet(B), B+avoparcin 10 ppm, B+nosiheptide 2.5 ppm and B+enramycin 5 ppm. Each treatment had five replications of 52 chicks each. In Experiment 2, two antibiotics (avoparcin and enramycin) were compared in $2{\times}2$ (antibiotics$\times$sex) factorial design. One thousand broiler chicks were reared in floor pens for 6 wk. Each of the 4 treatments had five replications of 50 chicks each. The results of Experiment 1 showed that antibiotic treatments (enramycin, avoparcin and nosiheptide) significantly(P <0.05) improved weight gain. Feed/gain ratio of avoparcin treatment and enramycin treatment tended to he lower than the control but they were not statistically significant. Dressing percentages were high in avoparcin and enramycin treatments compared to the control and nosiheptide treatment. The number of E. coli and Cl. perfringens in ileum and cecal contents were decreased by antibiotic treatments. Moisture contents in excreta of the birds were not significantly affected by the treatments. Leg abnormality and mortality were not significaniy different among treatments. In Experiment 2, weight gain was significantly (P<0.01) different between sexes, but not between antibiotic treatments. Significant effects of antibiotics (P<0.01), sex (P<0.01) and interaction (P<0.05) were shown in feed in take. Feed/gain ratio of avoparcin treatment was significantly (P<0.01) lower than that of enramycin treatment. leg abnormality and mortality were not significantly different among treatments but those in male broiler tended to be higher than in female broilers. It was concluded that nonsystemic antibiotics supplemented to the broiler diets suppress undesirable microorganisms and improve broiler performace in general and avoparcin was most effective in improving feed/gain ratio.
One of the biggest challenges for the animal feed industry in the coming years will be to meet the growing demand in animal protein in light of increased cost of feed ingredient as well as tougher restrictions on the use of antimicrobial growth promoters imposed by consumers and governments. A key focus area will be to maximise feed efficiency and minimise nutrient waste. It has been widely acknowledged that the composition of the intestinal microflora is closely related to intestinal health and performance of animals. Advanced microbial techniques have shown a close relationship between bacterial communities and their ability to modulate nutrient absorption and processing. In addition it has been recognised that modulating the immune response has significant impact on overall health as well as overall nutrient demand. Molecular techniques are a useful tool to gain an understanding of the impact of dietary interventions including the use of functional feed additives on specific changes in microbial communities or the immune system. Most these techniques however focus on the evaluation of large changes in bacterial compositions and often underestimate or neglect to recognise small changes in microbial diversity or behaviour changes without any measurable immune response. The key to understanding the relationship between specific nutritional intervention and the impact on health and performance lies in a deeper understanding of the impact of these nutrients on the expression of specific genes or specific metabolic pathways. The development of molecular tools as a result of developments in the field of Nutrigenomics has enabled researchers to study the effects of specific nutrients on the whole genome or in other words, the effect of thousands of genes simultaneously, and has opened a completely different avenue for nutritional research.
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