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Yoo, Sun Kyun,Gaugler, Randy,Brey, Christopher W. 한국산업미생물학회 2001 한국미생물·생명공학회지 Vol.29 No.2
공생 박테리아Photorhabdus sp. strain TF 균체량이 증가함에 따라서 병원성 선충 Heterorhabditis bacteriophora의 Infective Juveniles의 생산이 증가 되었다. 이 공생 박테리아의 성장 최적 조건은 각각 배양 온도 30℃ 그리고 초기 배지 pH 5.5 부터 7.3 사이에서 얻어졌다. 통기 상태에서 박테리아 균체의 생산과 성장이 촉진됨이 밝혀졌다. Photorhabdus sp. strain TF 박테리아의 성장 중 생산된 색소와 배양액의 pH의 변화는 박테리아의 성장 정도 나타내어 액체배양에서 병원성 선충 Heterorhabditis bacteriophora의 접종시기로 표시로서 이용할 수 있을 것이다. The yield of infective juveniles of Heterorhabditis bacteriophora (Tf strain) in vitro monoxenic liquid culture was improved significantly as the amount of symbiont biomass, Photorhabdus sp. strain Tf, increased. To investigate the influence of abiotic factors on the growth and biomass production of Photorhabdus sp. strain Tf, triplicate flask cultures were performed. The optinal temperature and medium pH for the growth of Photorhabdus sp. strain Tf were 30℃ and between pH 5.5~7.3, respectively. Aeration also improved greatly growth and yield of biomass of Photorhabdus sp. strain Tf. Photorhabdus sp. strain Tf in batch fermentation showed growth-associated pattern in terms of pigment production, and the pH of culture medium rose steadily until growth stopped during the fermentation. Both pigment production and culture pH rise would be useful parameters indicating a reliable growth of Photorhabdus sp. strain Tf.
An essential complementary role of NF-κB pathway to microbicidal oxidants in Drosophila gut immunity
Ryu, Ji-Hwan,Ha, Eun-Mi,Oh, Chun-Taek,Seol, Jae-Hong,Brey, Paul T,Jin, Ingnyol,Lee, Dong Gun,Kim, Jaesang,Lee, Daekee,Lee, Won-Jae Wiley (John WileySons) 2006 The EMBO journal Vol.25 No.15
<P>In the Drosophila gut, reactive oxygen species (ROS)-dependent immunity is critical to host survival. This is in contrast to the NF-kappaB pathway whose physiological function in the microbe-laden epithelia has yet to be convincingly demonstrated despite playing a critical role during systemic infections. We used a novel in vivo approach to reveal the physiological role of gut NF-kappaB/antimicrobial peptide (AMP) system, which has been 'masked' in the presence of the dominant intestinal ROS-dependent immunity. When fed with ROS-resistant microbes, NF-kappaB pathway mutant flies, but not wild-type flies, become highly susceptible to gut infection. This high lethality can be significantly reduced by either re-introducing Relish expression to Relish mutants or by constitutively expressing a single AMP to the NF-kappaB pathway mutants in the intestine. These results imply that the local 'NF-kappaB/AMP' system acts as an essential 'fail-safe' system, complementary to the ROS-dependent gut immunity, during gut infection with ROS-resistant pathogens. This system provides the Drosophila gut immunity the versatility necessary to manage sporadic invasion of virulent pathogens that somehow counteract or evade the ROS-dependent immunity.</P>
McQuilling, J.P.,Pareta, R.,Sivanandane, S.,Khanna, O.,Jiang, B.,Brey, E.M.,Orlando, G.,Farney, A.C.,Opara, E.C. Techno-Press 2014 Biomaterials and biomedical engineering Vol.1 No.1
We have previously described a new multilayer alginate microcapsule system, and the goals of the present study were to assess the in vitro function of islets encapsulated in its inner layer, and the angiogenic ability of FGF-1 delivered from the external layer in an omentum pouch. Following isolation and culture, islets were encapsulated in the inner core of microspheres ($500-600{\mu}m$ in diameter) with a semi-permeable poly-L-ornithine (PLO) membrane separating two alginate layers, and both unencapsulated and encapsulated islet function was assessed by a dynamic glucose perifusion. For angiogenesis experiments, one group of microcapsules without FGF-1 (control) and another (test) containing FGF-1 with heparin encapsulated in the external layer were made. One hundred microcapsules of each group were transplanted in Lewis rats (n = 5/group) and were retrieved after 14 days for assessment of angiogenesis. Glucose perifusion of unencapsulated and encapsulated islets resulted in similar stimulation indices. The release of FGF-1 resulted in increased vascular density compared to controls. In conclusion, islets encapsulated in the core of multilayer alginate microcapsules maintain functionality and the microcapsule's external layer is effective in delivery of FGF-1 to enhance graft neovascularization in a retrievable omentum pouch.
Han, S.-H.,Ryu, J.-H.,Oh, C.-T.,Nam, K.-B.,Nam, H.-J.,Jang, I.-H.,Brey, P. T.,Lee, W.-J. Blackwell Science Ltd 2004 Insect molecular biology Vol.13 No.3
<P>Abstract</P><p>The homeobox gene, <i>Caudal</i>, encodes the DNA-binding nuclear transcription factor that plays a crucial role during development and innate immune response. The <i>Drosophila</i> homologue of importin-7 (DIM-7), encoded by <i>moleskin</i>, was identified as a Caudal-interacting molecule during yeast two-hybrid screening. Both mutation of the minimal region of Caudal responsible for moleskin binding and RNA interference (RNAi) of <i>moleskin</i> dramatically inhibited the Caudal nuclear localization. Furthermore, Caudal-mediated constitutive expression of antifungal <i>Drosomycin</i> gene was severely affected in the <i>moleskin-</i>RNAi flies, showing a local <i>Drosomycin</i> expression pattern indistinguishable from that of the <i>Caudal-</i>RNAi flies. These <i>in vivo</i> data suggest that DIM-7 mediates Caudal nuclear localization, which is important for the proper Caudal function necessary for regulating innate immune genes in <i>Drosophila</i>.</p>
An Antioxidant System Required for Host Protection against Gut Infection in <i>Drosophila</i>
Ha, Eun-Mi,Oh, Chun-Taek,Ryu, Ji-Hwan,Bae, Yun-Soo,Kang, Sang-Won,Jang, In-hwan,Brey, Paul T.,Lee, Won-Jae Elsevier 2005 DEVELOPMENTAL CELL Vol.8 No.1
<P><B>Abstract</B></P><P>A fundamental question that applies to all organisms is how barrier epithelia efficiently manage continuous contact with microorganisms. Here, we show that in <I>Drosophila</I> an extracellular immune-regulated catalase (IRC) mediates a key host defense system that is needed during host-microbe interaction in the gastrointestinal tract. Strikingly, adult flies with severely reduced IRC expression show high mortality rates even after simple ingestion of microbe-contaminated foods. However, despite the central role that the NF-κB pathway plays in eliciting antimicrobial responses, NF-κB pathway mutant flies are totally resistant to such infections. These results imply that homeostasis of redox balance by IRC is one of the most critical factors affecting host survival during continuous host-microbe interaction in the gastrointestinal tract.</P>
Jang, In-Hwan,Chosa, Naoyuki,Kim, Sung-Hee,Nam, Hyuck-Jin,Lemaitre, Bruno,Ochiai, Masanori,Kambris, Zakaria,Brun, Sylvain,Hashimoto, Carl,Ashida, Masaaki,Brey, Paul T.,Lee, Won-Jae Elsevier 2006 Developmental cell Vol.10 No.1
<P><B>Summary</B></P><P>The Toll receptor was originally identified as an indispensable molecule for <I>Drosophila</I> embryonic development and subsequently as an essential component of innate immunity from insects to humans. Although in <I>Drosophila</I> the Easter protease processes the pro-Spätzle protein to generate the Toll ligand during development, the identification of the protease responsible for pro-Spätzle processing during the immune response has remained elusive for a decade. Here, we report a protease, called Spätzle-processing enzyme (SPE), required for Toll-dependent antimicrobial response. Flies with reduced SPE expression show no noticeable pro-Spätzle processing and become highly susceptible to microbial infection. Furthermore, activated SPE can rescue ventral and lateral development in embryos lacking Easter, showing the functional homology between SPE and Easter. These results imply that a single ligand/receptor-mediated signaling event can be utilized for different biological processes, such as immunity and development, by recruiting similar ligand-processing proteases with distinct activation modes.</P>