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( Jiyoung Shim ),( Langley Williams ),( Dohyun Kim ),( Kisung Ko ),( Moon-soo Kim ) 한국미생물 · 생명공학회 2021 Journal of microbiology and biotechnology Vol.31 No.9
Micro-scale magnetic beads are widely used for isolation of proteins, DNA, and cells, leading to the development of in vitro diagnostics. Efficient isolation of target biomolecules is one of the keys to developing a simple and rapid point-of-care diagnostic. A zinc finger protein (ZFP) is a doublestranded (ds) DNA-binding domain, providing a useful scaffold for direct reading of the sequence information. Here, we utilized two engineered ZFPs (Stx2-268 and SEB-435) to detect the Shiga toxin (stx2) gene and the staphylococcal enterotoxin B (seb) gene present in foodborne pathogens, Escherichia coli O157 and Staphylococcus aureus, respectively. Engineered ZFPs are immobilized on a paramagnetic bead as a detection platform to efficiently isolate the target dsDNA-ZFP bound complex. The small paramagnetic beads provide a high surface area to volume ratio, allowing more ZFPs to be immobilized on the beads, which leads to increased target DNA detection. The fluorescence signal was measured upon ZFP binding to fluorophore-labeled target dsDNA. In this study, our system provided a detection limit of ≤ 60 fmol and demonstrated high specificity with multiplexing capability, suggesting a potential for development into a simple and reliable diagnostic for detecting multiple pathogens without target amplification.
도명술,정헌순,최봉혁,Leif Hunter,Stuart Langley,Laszlo Pazmany,Paul Trayhurn 연세대학교의과대학 2006 Yonsei medical journal Vol.47 No.5
We report here the use of human inflammation arrays to study the inflammatory gene expression profile of TNF-α- treated human SGBS adipocytes. Human preadipocytes (SGBS) were induced to differentiate in primary culture, and adipocyte differentiation was confirmed, using Oil Red O staining. We treated the differentiated adipocytes with TNF-α, and RNA from differentiated adipocytes with or without TNF-α treatment was hybridized to MWG human inflammation arrays to compare expression profiles. Eleven genes were up- or down-regulated in TNF-α-treated adipocytes. As revealed by array analysis, among 6 up-regulated genes, only eotaxin-1, monocyte chemoattractant protein-1 (MCP-1), and vascular cell adhesion molecule 1 isoform a precursor (VCAM1) were confirmed by real-time polymerase chain reaction (PCR). Similarly, among 5 down-regulated genes, only IL-1 family member 5 (IL1F5), a disintegrin and metalloprotease with thrombospondin motifs-1 preproprotein (ADAMTS1), fibronectin 1 isoform 1 preprotein (FN1), and matrix metalloproteinase 15 preprotein (MMP15) were confirmed by real-time PCR. There was a substantial increase (50-fold) in eotaxin-1 in response to TNF-α. Taken together, we have identified several inflammatory molecules expressed in SGBS adipocytes and discovered molecular factors explaining the relationship between obesity and atherosclerosis, focusing on inflammatory cytokines expressed in the TNF-α-treated SGBS cells. Further investigation into the role of these up- or down-regulated cytokine genes during the pathological processes leading to the development of atherosclerosis is warranted.
Lee, Seung-Hoon,Megonigal, Patrick J.,Langley, Adam J.,Kang, Hojeong Elsevier 2017 Applied soil ecology Vol.117 No.-
<P><B>Abstract</B></P> <P>With unique and important characteristics, salt marsh ecosystems are expected to be affected by elevated CO<SUB>2</SUB> and N enrichment. Although various studies have assessed the effects of those changes on the vegetation of salt marshes, little information is available about their impact on microbes. In this study, we comprehensively analyzed the microbial community structure of soils responding to elevated CO<SUB>2</SUB> (eCO<SUB>2</SUB>) and N addition (N) over 3 years in salt marsh ecosystem. We employed pyrosequencing, T-RFLP analysis, and quantitative PCR to study the bacterial, archaeal, and fungal communities. The overall results indicated that 1) eCO<SUB>2</SUB> and N affected the microbial abundance but not community structure significantly in salt marsh system, 2) due to their different ecophysiology, the responses of the three different microbial communities to eCO<SUB>2</SUB> and/or N addition varied with each group, 3) the composition (C3/C4 or diversity) of the plant community was important in structuring the microbial community of salt marsh ecosystems, which generally have low plant diversity, 4) the archaeal and fungal communities responded more strongly to eCO<SUB>2</SUB> and/or N addition than the bacterial community. This study represents the first comprehensive report of the effects of eCO<SUB>2</SUB> and/or N addition on the diverse microbial community structures of tidal marsh systems. It suggests that single or combined effect of eCO<SUB>2</SUB> and N on microbial abundance in salt marsh was obvious, and that the key groups playing an important role in the biogeochemical process can be shifted.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Elevated CO<SUB>2</SUB> and/or N affected the microbial abundance but not structure significantly. </LI> <LI> The responses of bacterial, archaeal, and fungal communities varied with each group. </LI> <LI> The plant C3/C4 composition was important in structuring the microbial communities. </LI> <LI> The archaeal and fungal communities responded to eCO<SUB>2</SUB> and/or N more strongly. </LI> </UL> </P>
Yu, Hyun-Kyung,Lee, Ho-Jeong,Yun, Seok-Joong,Lee, Sun-Joo,Langley, Robert R.,Yoon, Yeup,Yi, Lee S.H.,Bae, Duk-Soo,Kim, Jang-Seong,Kim, Sun Jin Neoplasia Press 2014 Translational oncology Vol.7 No.3
<P><I>INTRODUCTION:</I> The present study compared the effect of combination therapy using human apolipoprotein(a) kringle V (rhLK8) to conventional chemotherapy with paclitaxel for human ovarian carcinoma producing high or low levels of vascular endothelial growth factor (VEGF). <I>MATERIALS AND METHODS:</I> Human ovarian carcinoma cells producing high (SKOV3ip1) or low (HeyA8) levels of VEGF were implanted into the peritoneal cavity of female nude mice. Seven days later, mice were randomized into four groups: control (vehicle), paclitaxel [5 mg/kg, weekly intraperitoneal (i.p.) injection], rhLK8 (50 mg/kg, daily i.p. injection), or the combination of paclitaxel and rhLK8. Mice were treated for 4 weeks and examined by necropsy. <I>RESULTS:</I> In mice implanted with SKOV3ip1 cells, rhLK8 treatment had no significant effect on tumor incidence or the volume of ascites but induced a significant decrease in tumor weight compared with control mice. Paclitaxel significantly reduced tumor weight and ascites volume, and combination treatment with paclitaxel and rhLK8 had an additive therapeutic effect. Similarly, in HeyA8 mice, the effect of combination treatment on tumor weight and tumor incidence was statistically significantly greater than that of paclitaxel or rhLK8 alone. Immunohistochemical analysis showed a significant decrease in microvessel density and a marked increase of apoptosis in tumor and tumor-associated endothelial cells in response to combination treatment with paclitaxel and rhLK8. <I>CONCLUSION:</I> Collectively, these results suggest that antiangiogenic therapy with rhLK8 in combination with taxane-based conventional chemotherapy could be effective for the treatment of ovarian carcinomas, regardless of VEGF status.</P>
McFall, Haley,Sarabu, Sandeep,Shankar, Vijaykumar,Bandari, Suresh,Murthy, S. Narasimha,Kolter, Karl,Langley, Nigel,Kim, Dong Wuk,Repka, Michael A. Elsevier 2019 International journal of pharmaceutics Vol.554 No.-
<P><B>Abstract</B></P> <P>The objective of this study was to formulate aripiprazole (ARI)-loaded pH-modulated solid dispersions (SD) to enhance solubility, dissolution, and bioavailability via hot-melt extrusion (HME) technology. Kollidon® 12 PF (PVP) and succinic acid (SA) were selected after solubility screenings of various polymers and acidifiers. Several formulations, varying in screw speed and drug/polymer/acidifier ratios, were extruded using an 11 mm twin-screw extruder and were investigated for the effect of these variables. Scanning electron microscopy (SEM), differential scanning calorimetry (DSC), and X-ray diffraction (XRD) were used to perform solid-state characterizations of the pure drug and extrudates. The aqueous solubility and dissolution were evaluated for the pure drug and milled extrudates. Among the prepared formulations, N6 was chosen for <I>in vivo</I> absorption studies. Solid-state characterization demonstrated the transformation of the crystalline ARI to an amorphous state in the formulations. Each formulation showed increased solubility and dissolution compared to the drug powder. The oral bioavailability (C<SUB>max</SUB> and AUC<SUB>0–12</SUB>) of N6 was significantly improved when compared to the pure ARI. This novel study not only discusses the incorporation of acidifiers in SDs but also the preparation of SDs using HME technology as effective techniques to improve drug release and bioavailability.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>
EphA2 immunoconjugate as molecularly targeted chemotherapy for ovarian carcinoma.
Lee, Jeong-Won,Han, Hee Dong,Shahzad, Mian M K,Kim, Seung Wook,Mangala, Lingegowda S,Nick, Alpa M,Lu, Chunhua,Langley, Robert R,Schmandt, Rosemarie,Kim, Hye-Sun,Mao, Shenlan,Gooya, John,Fazenbaker, Ch U.S. Dept. of Health, Education, and Welfare, Publ 2009 Journal of the National Cancer Institute Vol.101 No.17
<P>EphA2 is overexpressed in many types of human cancer but is absent or expressed at low levels in normal epithelial tissues. We investigated whether a novel immunoconjugate containing an anti-EphA2 monoclonal antibody (1C1) linked to a chemotherapeutic agent (monomethyl auristatin phenylalanine [MMAF]) through a noncleavable linker maleimidocaproyl (mc) had antitumor activity against ovarian cancer cell lines and tumor models.</P>