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Flores-Fernandez, Carol N.,Chavez-Hidalgo, Elizabeth,Santos, Marco,Zavaleta, Amparo I.,Arahal, David R. The Korean Society for Microbiology and Biotechnol 2019 한국미생물·생명공학회지 Vol.47 No.3
All Idiomarina species are isolated from saline environments; microorganisms in such extreme habitats develop metabolic adaptations and can produce compounds such as proteases with an industrial potential. ARDRA and 16S rRNA gene sequencing are established methods for performing phylogenetic analysis and taxonomic identification. However, 16S-23S ITS is more variable than the 16S rRNA gene within a genus, and is therefore, used as a marker to achieve a more precise identification. In this study, ten protease producing Idiomarina strains isolated from the Peruvian salterns were characterized using biochemical and molecular methods to determine their bacterial diversity and industrial potential. In addition, comparison between the length and nucleotide sequences of a 16S-23S ITS region allowed us to assess the inter and intraspecies variability. Based on the 16S rRNA gene, two species of Idiomarina were identified (I. zobellii and I. fontislapidosi). However, biochemical tests revealed that there were differences between the strains of the same species. Moreover, it was found that the ITS contains two tRNA genes, $tRNA^{Ile(GAT)}$ and $tRNA^{Ala(TGC)}$, which are separated by an ISR of a variable size between strains of I. zobellii. In one strain of I. zobellii (PM21), we found nonconserved nucleotides that were previously not reported in the $tRNA^{Ala}$ gene sequences of Idiomarina spp. Thus, based on the biochemical and molecular characteristics, we can conclude that protease producing Idiomarina strains have industrial potential; only two I. zobellii strains (PM48 and PM72) exhibited the same properties. The differences between the other strains could be explained by the presence of subspecies.
Interleukin-17A production in Riemerella anatipestifer infection is independent to IL-23
Flores Rochelle(Flores Rochelle ),Fernandez Cherry(Fernandez Cherry ),Afrin Fahmida(Afrin Fahmida ),Cammayo Paula Leona(Cammayo Paula Leona ),Roy Anindita(Roy Anindita ),Min Wongi(Min Wongi ) 한국예방수의학회 2019 한국예방수의학회 학술대회자료집 Vol.2018 No.-
Fernandez, Cherry P.,Afrin, Fahmida,Flores, Rochelle A.,Kim, Woo H.,Jeong, Jipseol,Kim, Suk,Lillehoj, Hyun S.,Min, Wongi Elsevier 2018 Molecular immunology Vol.95 No.-
<P>As the dysregulation of IL-17 is implicated in the pathogenesis of various autoimmune and inflammatory diseases, the suppression of IL-17 production by Th2 cytokines could alleviate the development of these diseases. Previously, we confirmed that inflammatory cytokines including IL-17A are strongly associated with R. anatipestifer infection, which is one of the most important bacterial pathogens in the duck industry. Here, we found that IL-4 treatment downregulated the expression of IL-17A and IL-17F transcripts in splenic lymphocytes stimulated with R. anatipestifer. Moreover, duck IL-4 (duIL-4) treatment in R. anatipestifer-stimulated lymphocytes suppressed the expression of IL-23p19 and IL-12p40 transcripts compared to untreated and stimulated lymphocytes. Conversely, dulL-4 increased levels of IFN-gamma and IL-10. We identified a full-length dulL-4 cDNA encoding 136 amino acids from ConA-activated splenic lymphocytes that shares 49.3-50% amino acid sequence identity with chicken and quail IL-4 and 21-29.7% with mammalian and piscine homologues. Low or moderate levels of dulL-4 transcript were observed in healthy tissues, including the spleen, bursa, and thymus, whereas dulL-4 expression was higher in the kidney and lung. Levels of dulL-4 were generally upregulated in mitogen-activated splenic lymphocytes but lower in the liver and spleen of R. anatipestifer-infected ducks compared to those of infected chickens. Recombinant dulL-4 promoted nitric oxide synthesis in duck macrophages stimulated by It anatipestifer compared to untreated and stimulated control macrophages. These results demonstrate that IL-4 is an important Th2 cytokine that inhibits inflammatory responses in splenic lymphocytes stimulated with R. anatipestifer.</P>
Fernandez, Cherry P.,Afrin, Fahmida,Flores, Rochelle A.,Kim, Woo H.,Jeong, Jipseol,Kim, Suk,Chang, Hong H.,Lillehoj, Hyun S.,Min, Wongi PERGAMON 2017 DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY Vol. No.
<P><B>Abstract</B></P> <P> <I>Riemerella anatipestifer,</I> an important infectious bacterium affecting the duck industry, has 5–75% mortality, depending on strain virulence. We previously demonstrated that proinflammatory cytokines are involved in inflammation during, and regulating susceptibility to, <I>R. anatipestifer</I> infection. We investigated the effects of the anti-inflammatory compound berberine in duck splenic lymphocytes stimulated with killed <I>R. anatipestifer,</I> and in <I>R. anatipestifer-</I>infected ducks. IL-17A, IL-17F, and IL-1β transcripts were downregulated, and IFN-γ and IL-10 transcripts enhanced, in berberine-treated stimulated splenic lymphocytes, compared to stimulated untreated splenic lymphocytes. Similarly, IL-17A, IL-17F, IL-6, and IL-1β expressions were significantly reduced, and IFN-γ and IL-10 expressions significantly upregulated, in spleens and livers of <I>R. anatipestifer-</I>infected berberine-treated ducks, compared to infected untreated birds. Moreover, infected and treated birds showed increased survival rates and significantly decreased bacterial burdens compared to infected untreated birds, confirming that inflammatory cytokines are strongly associated with <I>R. anatipestifer</I> infection in ducks.</P> <P><B>Highlights</B></P> <P> <UL> <LI> <I>R. anatipestifer</I> infection is ameliorated by inflammatory cytokine suppression. </LI> <LI> Expression levels of IL-17A, IL-17F, IL-1β or IL-6 mRNA were significantly reduced by berberine treatment. </LI> <LI> Expression of anti-inflammatory cytokine IL-10 was enhanced <I>in vitro</I> and <I>in vivo.</I> </LI> <LI> Berberine-treated infected ducks had lower mortality rate and bacterial load. </LI> </UL> </P>
Rochelle Alipio Flores,Binh Thanh Nguyen,Paula Leona Tayamen Cammayo,Cherry Pambid Fernandez-Colorado,Anindita Roy,Suk Kim,Woohyun Kim,Wongi Min 대한수의학회 2022 Journal of Veterinary Science Vol.23 No.6
Three parameters, body weight gain (BWG), intestinal lesion score (LS) and fecal oocyst shedding, were compared in broilers infected with major parasitic species; Eimeria acervulina, E. maxima, and E. tenella. First, two- and three-week-old chickens with Eimeria infection showed LS of approximately 3, but two-week-old chickens were more correlated with BWG. Second, significant differences in BWG were observed between male and female broilers challenged with Eimeria. Finally, E. maxima-infected broilers among three Eimeria species showed a higher relationship between BWG and LS, suggesting three considerations such as genders, age and Eimeria species for Eimeria experiments.