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Quercetin Induces Mitochondrial Biogenesis through Activation of HO-1 in HepG2 Cells
Rayamajhi, Nabin,Kim, Seul-Ki,Go, Hiroe,Joe, Yeonsoo,Callaway, Zak,Kang, Jae-Gu,Ryter, Stefan W.,Chung, Hun Taeg Hindawi Publishing Corporation 2013 Oxidative medicine and cellular longevity Vol.2013 No.-
<P>The regeneration of mitochondria by regulated biogenesis plays an important homeostatic role in cells and tissues and furthermore may provide an adaptive mechanism in certain diseases such as sepsis. The heme oxygenase (HO-1)/carbon monoxide (CO) system is an inducible cytoprotective mechanism in mammalian cells. Natural antioxidants can provide therapeutic benefit, in part, by inducing the HO-1/CO system. This study focused on the mechanism by which the natural antioxidant quercetin can induce mitochondrial biogenesis in HepG2 cells. We found that quercetin treatment induced expression of mitochondrial biogenesis activators (PGC-1<I><I>α</I></I>, NRF-1, TFAM), mitochondrial DNA (mtDNA), and proteins (COX IV) in HepG2 cells. The HO inhibitor SnPP and the CO scavenger hemoglobin reversed the effects of quercetin on mitochondrial biogenesis in HepG2 cells. The stimulatory effects of quercetin on mitochondrial biogenesis could be recapitulated <I>in vivo</I> in liver tissue and antagonized by SnPP. Finally, quercetin conferred an anti-inflammatory effect in the liver of mice treated with LPS and prevented impairment of mitochondrial biogenesis by LPS <I>in vivo</I>. These salutary effects of quercetin <I>in vivo</I> were also antagonized by SnPP. Thus, our results suggest that quercetin enhances mitochondrial biogenesis mainly via the HO-1/CO system <I>in vitro</I> and <I>in vivo</I>. The beneficial effects of quercetin may provide a therapeutic basis in inflammatory diseases and sepsis.</P>
Rayamajhi, Nabin,Cha, Seung Bin,Kang, Mi Lan,Lee, Su In,Lee, Hee Soo,Yoo, Han Sang American Society for Microbiology 2009 Applied and environmental microbiology Vol.75 No.17
<B>ABSTRACT</B><P>Florfenicol resistance was analyzed in 230 enteric pig isolates collected between 1998 and 2006. PCR, plasmid profiling, Southern blot hybridization, and a mixed-broth conjugation assay suggested the intra- and interspecies plasmid-mediated transfer of florfenicol resistance among the isolates that exhibited MICs for florfenicol between 4 to 128 mg/liter.</P>
Rayamajhi, Nabin,Jung, Byeong Yeal,Cha, Seung Bin,Shin, Min Kyung,Kim, Aeran,Kang, Min Su,Lee, Kang Mu,Yoo, Han Sang American Society for Microbiology 2010 Applied and environmental microbiology Vol.76 No.14
<B>ABSTRACT</B><P>Fifteen nonrepetitive ampicillin-resistant <I>Salmonella</I> spp. were identified among 91 <I>Salmonella</I> sp. isolates during nationwide surveillance of <I>Salmonella</I> in waste from 131 chicken farms during 2006 and 2007. Additional phenotyping and genetic characterization of these 15 isolates by using indicator cephalosporins demonstrated that resistance to ampicillin and reduced susceptibility to cefoxitin in three isolates was caused by TEM-1 and DHA-1 β-lactamases. Plasmid profiling and Southern blot analysis of these three DHA-1-positive <I>Salmonella</I> serovar Indiana isolates and previously reported unrelated clinical isolates of DHA-1-positive <I>Salmonella</I> serovar Montevideo, <I>Klebsiella pneumoniae</I>, and <I>Escherichia coli</I> from humans and swine indicated the involvement of the large-size plasmid. Restriction enzyme digestion of the plasmids from the transconjugants showed variable restriction patterns except for the two <I>S</I><I>almonella</I> serovar Indiana isolates identified in this study. To the best of our knowledge, this is the first report of the presence of the DHA-1 gene among <I>Salmonella</I> spp. of animal origin.</P>
Isolation, Characterization, and Evaluation of Wild Isolates of Lactobacillus reuteri from Pig Feces
Deog Yong Lee,Yeon-Soo Seo,Nabin Rayamajhi,강미란,Su In Lee,유한상 한국미생물학회 2009 The journal of microbiology Vol.47 No.6
Lactic acid bacteria (LAB) are a well-used probiotics for health improvements in both humans and animals. Despite of several benefits, non-host-specific LAB showed poor probiotics effects due to difficulty in colonization and competition with normal flora. Therefore, the feasibility of porcine LAB isolates was evaluated as a probiotics. Ten of 49 Lactobacillus spp. isolates harbored 2~10 kb plasmid DNA. Seven strains were selected based on the safety test, such as hemolytic activity, ammonia, indole, and phenylalanine production. After safety test, five strains were selected again by several tests, such as epithelial adherence, antimicrobial activity, tolerance against acid, bile, heat, and cold-drying, and production of acid and hydrogen peroxide. Then, enzyme profiles (ZYM test) and antibiotics resistance were analyzed for further characterization. Five Lactobacillus reuteri isolates from pig feces were selected by safety and functional tests. The plasmid DNA which was able to develop vector system was detected in the isolates. Together with these approaches, pig-specific Lactobacillus spp. originated from pigs were selected. These strains may be useful tools to develop oral delivery system.
Lee, Deog-Yong,Kang, Sang-Gyun,Rayamajhi, Nabin,Kang, Milan,Yoo, Han Sang The Korean Society of Veterinary Science 2008 大韓獸醫學會誌 Vol.48 No.4
The genus Lactobacillus is the largest of the genera included in lactic acid bacteria and is associated with mucosal membranes of human and animal. Only a few Lactobacillus plasmid-encoded functions have been discovered and used. In this study, a novel plasmid (pILR091) was isolated from a wild L. reuteri isolated from pig and described the characteristics of its replicons, genetic organization, and relationship with other plasmids. After digestion of the plasmid, pILR091, with SalI, plasmid DNA was cloned into the pQE-30Xa vector and sequenced. The complete sequence was confirmed by the sequencing of PCR products and analyzed with the Genbank database. The isolate copy number and stability were determined by quantitative-PCR. The complete sequence of L. reuteri contained 7,185 nucleotides with 39% G-C content and one cut site by two enzymes, SalI and HindIII. The similar ori sequence of the pC194- rolling circle replication family (TTTATATTGAT) was located 63 bp upstream of the protein replication sequence, ORF 1. Total of five ORFs was identified and the coding sequence represented 4,966 nucleotides (70.4%). ORF1 of pILR091 had a low similarity with the sequence of pTE44. Other ORFs also showed low homology and E-values. The average G-C content of pILR091 was 39%, similar with that of genomic DNA. The copy number of pILR091 was determined at approximately 24 to 25 molecules per genomic DNA. These results suggested that pILR091 might be a good candidate to construct a new vector, which could be used for cloning and expression of foreign genes in lactobacilli.
Bacteriophage의 Salmonella enterica serovar Enteritidis에 대한 in vitro 및 in vivo 효능 평가
차승빈,이원정,신민경,노유미,정명환,명길선,안영태,허철성,유한상,Cha, Seung-Bin,Rayamajhi, Nabin,Lee, Won-Jung,Shin, Min-Kyoung,Roh, Yu-Mi,Jung, Myung-Hwan,Myoung, Kil-Sun,Ahn, Young-Tae,Huh, Chul-Sung,Yoo, Han Sang 대한수의학회 2010 大韓獸醫學會誌 Vol.50 No.3
Salmonella (S.) Enterica infection ranks among the most common food borne bacterial infections worldwide. Although there are six subspecies of S. Enterica, the vast majority of human and animal infections are caused by strains belonging to subspecies 1 serovar Typhimurium and Enteritidis. Recent reports on antibiotic resistance of Salmonella spp. are rising steadily. The increasing problem of antibiotic resistance has rekindled interest in bacteriophage to therapy. Therefore, we investigated the efficacy of bacteriophage in S. enterica serovar Enteritidis infected mice and pigs by measuring of body condition, body weight, bacterial colonization and weight of organs based on the in vitro analysis. In vitro experiment, phage cultured with S. Enteritidis showed clear lysis pattern, the plaque forming unit (PFU) of our phage culture was $1.5{\times}10^{11}PFU/mL$, and phage showed its maximum activity at 4 h post inoculation. In mouse experiment, there was no significant difference among experimental groups in the general body conditions and body weight of mice. However, there was difference in weight of liver and spleen depending on the experimental group (p < 0.05). The weight of liver and spleen were reduced by the phage treatment. Also bacterial colonization in spleen and liver were significantly reduced by the phage treatment. In pig experiment, the general body conditions and body temperature exhibited not much difference among the pigs except few pigs in group 3 which showed poor body conditions. From the feces in each group, we could isolate the S. Enteritidis only from group 3. Bacterial enrichment culture was necessary for isolating the bacteria from 5 dpi and 10 dpi, however direct isolation was possible from 15 dpi feces. In phage treated group, postmortem lesion was better than non-phage treated group. Recently, antibiotic resistance concerns on the food-borne bacterial pathogens have been increasing because of the wide spread of the antibiotics resistance genes. This concern is widely transmitted to the human related public health. As one of the alternative treatments on the bacterial pathogens, attempt using phages have been made to control the bacterial diseases. The positive possibility of the trail using phage was observed to control the S. enterica serovar Enteritidis in this study even though the further analysis has been remained.