Development of a novel peptide microarray for large-scale epitope mapping of food allergens

Lin, Jing,Bardina, Ludmilla,Shreffler, Wayne G.,Andreae, Doerthe A.,Ge, Yongchao,Wang, Julie,Bruni, Francesca M.,Fu, Zhiyan,Han, Youngshin,Sampson, Hugh A. Elsevier 2009 The Journal of allergy and clinical immunology Vol.124 No.2

<P><B>Background</B></P><P>The peptide microarray is a novel assay that facilitates high-throughput screening of peptides with a small quantity of sample.</P><P><B>Objective</B></P><P>We sought to use overlapping peptides of milk allergenic proteins as a model system to establish a reliable and sensitive peptide microarray-based immunoassay for large-scale epitope mapping of food allergens.</P><P><B>Methods</B></P><P>A milk peptide microarray was developed by using commercially synthesized peptides (20-mers, 3 offset) covering the primary sequences of α<SUB>s1</SUB>-casein, α<SUB>s2</SUB>-casein, β-casein, κ-casein, and β-lactoglobulin. Conditions for printing and immunolabeling were optimized using a serum pool of 5 patients with milk allergy. Reproducibility of the milk peptide microarray was evaluated using replicate arrays immunolabeled with the serum pool, whereas specificity and sensitivity were assessed by using serial dilution of the serum pool and a peptide inhibition assay.</P><P><B>Results</B></P><P>Our results show that epitopes identified by the peptide microarray were mostly consistent with those identified previously by SPOT membrane technology, but with specific binding to a few newly identified epitopes of milk allergens. Data from replicate arrays were reproducible (<I>r</I> ≥ 0.92) regardless of printing lots, immunolabeling, and serum pool batches. Using the serially diluted serum pool, we confirmed that IgE antibody binding detected in the array was specific. Peptide inhibition of IgE binding to the same peptide and overlapping peptides further confirmed the specificity of the array.</P><P><B>Conclusion</B></P><P>A reliable peptide microarray was established for large-scale IgE epitope mapping of milk allergens, and this robust technology could be applied for epitope mapping of other food allergens.</P>

‘I Only Hate Broccoli’: The Library as Place in 21st Century America

Wayne A. Wiegand 한국문헌정보학회 2010 한국문헌정보학회지 Vol.44 No.4

By taking a bottom-up “library in the life of the user” perspective rather than a top-down “user in the life of the library” perspective, this paper uses anecdotal evidence from the past and near present to examine the multiple roles the U.S. public library plays and has played as public space in the everyday lives of its patrons. By harnessing “public sphere ” theory discussed in Jürgen Habermas’s THE STRUCTURAL TRANSFORMATION OF THE PUBLIC SPHERE (1989) and by the examining the rich literatures on civic life and institutions that have evolved from it, the author argues that Library and Information Studies discourse has to expand its scope to include research and analysis of “library as place” from a user’s perspective if it hopes to develop a deeper understanding of what the public library does for means to members of the communities in which they reside.

유방암에 대한 제니스타인의 효과 : 화학적 예방요법, 생체작용 기작과 생체 이용법 Chemoprevention, In Vivo Mechanisms of Action, and Bioavailability

Lamartiniere, Coral A.,Fritz, Wayne A. 한국콩연구회 1998 韓國콩硏究會誌 Vol.15 No.2

Genistein은 대두 이소플라본의 주요 성분이다. 우리는 실험용 쥐를 이용하여 암 예방에 미치는 효과와 독성, 그 작용 기작과 생체 이용성에 미치는 Genistein의 잠재적 가능성을 밝히는데 주력하여 연구하였다. 임신부터 분만까지 21일 동안 실험용 쥐에게 식이를 통하여 Genistein을 공급시(25㎎, 250㎎ Genistein/㎏식이) 성체의 암컷쥐에게 화학적으로 유발된 유선종양의 발생이 genistein용량에 따라 변화한다는 결과를 얻었다. Whole mount 기법을 이용한 유선의 분석 결과 21일령과 50일령 암컷 쥐의 TEB(terminal end bud)의 수는 적었으며, 50일령의 terminal duct는 미분화된 상태로 발암 감수성이 높았다. 일일 식이 1㎏당 genistein 250㎎을 섭취한 어미쥐의 분만 7일차 에 재취한 혈청과 유즙의 총 genistein 농도는 각각 418±198 pmol/㎖ 이었다. 생 후 7일령의 새끼쥐의 위에서 채취한 유즙과 혈청 및 유선에서의 총 genistein 함량은 각각 4439± 1109pmol/㎖, 726pmol/㎖, 440± 129pmol/㎖ 이었으며, 생 후 21일령 쥐의 혈청과 유선의 총 genistein 수준은 1,810pmol/㎖과 370± 36pmol/㎖ 이었다. 우리의 연구는 genistein이 암컷의 유선과 수컷의 전립선에서 에스트로겐 수용체와 EGF 수용체와 반응하여 작용한다는 것을 보였다. 태생기 동안의 식이 중의 genistein은 0세대 암컷의 생식력에 유의적인 변화를 미치지 못하였고, 1세대 수컷과 암컷의 산자수, 질구개관, 성주기 기간, 난포 성숙에도 유의적인 영향을 끼치지 않았다. 조직형태학적 평가에 있어서도 태생기의 genistein 노출은 암컷의 생식기계에 어떠한 변화도 초래하지 않았다. 따라서 우리는 이러한 실험 결과에 의해 genistein은 10-9 물 농도 수준에서 독성이 없거나 극히 적으며 여러 기작에 의해 작용할 수 있다는 결론에 도달하였다. Cancer prevention rather than therapy is the desired future of cancer health care. The identification of dietary factors that can contribute to the prevention of cancer would be optimum. The primary cancers of men and women are breast and prostate, respectively. The incidence of these cancers are especially high in the Western World. Asians eating a diet high in soy products have lower incidences of breast and malignant prostate cancers than Americans (1-4). Another interesting fact is that of Asians emigrating to the U. S., it is the second generation, not the first generation, that loses the protection from eating soy (5). This has prompted us to hypothesize that a component of soy, early in life, would confer a $quot;programming$quot; effect on the breast to render a permanent protection against this cancer. Programming is defined as developmental alterations occurring during early critical periods of life that result in permanent manifestations, even in the absence of the original effector in adulthood.

Effect of Technetium-99 sources on its retention in low activity waste glass

Luksic, Steven A.,Kim, Dong-Sang,Um, Wooyong,Wang, Guohui,Schweiger, Michael J.,Soderquist, Chuck Z.,Lukens, Wayne,Kruger, Albert A. Elsevier 2018 JOURNAL OF NUCLEAR MATERIALS Vol.503 No.-

<P><B>Abstract</B></P> <P>Small-scale crucible melting tests on simulated waste glass were performed with technetium-99 (Tc-99) introduced as different species in a representative low activity waste simulant. The glass saw an increase in Tc-99 retention when TcO<SUB>2</SUB>∙2H<SUB>2</SUB>O and various Tc-minerals containing reduced tetravalent Tc were used compared to tests in which pertechnetate with heptavalent Tc was used. We postulate that the increase of Tc retention is likely caused by different reaction paths for Tc incorporation into glass during early stages of melting, rather than the low volatility of reduced tetravalent Tc compounds, which has been a generally accepted idea. Additional studies are needed to clarify the exact mechanisms relevant to the effect of reduced Tc compounds on Tc incorporation into or volatilization from the glass melt.</P>

Reduction and Simultaneous Removal of ⁹⁹Tc and Cr by Fe(OH)₂(s) Mineral Transformation

Saslow, Sarah A.,Um, Wooyong,Pearce, Carolyn I.,Engelhard, Mark H.,Bowden, Mark E.,Lukens, Wayne,Leavy, Ian I.,Riley, Brian J.,Kim, Dong-Sang,Schweiger, Michael J.,Kruger, Albert A. American Chemical Society 2017 Environmental science & technology Vol.51 No.15

<P>Technetium (Tc) remains a priority remediation concern due to persistent challenges, including mobilization due to rapid reoxidation of immobilized Tc, and competing comingled contaminants, e.g., Cr(VI), that inhibit Tc(VII) reduction and incorporation into stable mineral phases. Here Fe(OH)(2)(s) is investigated as a comprehensive solution for overcoming these challenges, by serving as both the reductant, (Fe(II)), and the immobilization agent to form Tc-incorporated magnetite (Fe3O4). Trace metal analysis suggests removal of Tc(VII) and Cr(VI) from solution occurs simultaneously; however, complete removal and reduction of Cr(VI) is achieved earlier than the removal/reduction of comingled Tc(VII). Bulk oxidation state analysis of the final magnetite solid phase by XANES shows that the majority of Tc is Tc(IV), which is corroborated by XPS measurements. Furthermore, EXAFS results show successful, albeit partial, Tc(IV) incorporation into magnetite octahedral sites. Cr XPS analysis indicates reduction to Cr(III) and the formation of a Cr-incorporated spinel, Cr2O3, and Cr(OH)(3) phases. Spinel (modeled as Fe3O4), goethite (alpha-FeOOH), and feroxyhyte (delta-FeOOH) are detected in all reacted final solid phase samples analyzed by XRD. Incorporation of Tc(IV) has little effect on the spinel lattice structure. Reaction of Fe(OH)(2)(s) in the presence of Cr(III) results in the formation of a spinel phase that is a solid solution between magnetite (Fe3O4) and chromite (FeCr2O4).</P>

Determination of Branched-Chain α-Keto Acid Dehydrogenase Activity in Rat Tissues

Kim, Hyun-Sook,Johnson, Wayne A. Korean Society for Biochemistry and Molecular Biol 1995 Journal of biochemistry and molecular biology Vol.28 No.1

The branched-chain ${\alpha}$-keto acid dehydrogenase (BCKAD) complex is a rate limiting enzyme which catalyzes the oxidative decarboxylation of branched-chain ${\alpha}$-keto acids. Numerous studies have suggested that BCKAD is subject to covalent modification in vitro via phosphorylation and dephosphorylation, which are catalyzed by a specific kinase and phosphatase, respectively. The biggest difficulty in the assay of BCKAD activity is to arrest the interconversion between the active and inactive forms. BCKAD activity was determined from fresh rat heart and liver tissues using homogenizing and assay buffers containing inhibitors of phosphatase and kinase. The results suggest that a radiochemical assay using ${\alpha}$-keto[1-$^{14}C$]-isovalerate as a substrate for the enzyme can be applied as a reliable method to determine in vitro enzyme activity with arrested interconversion between the active and inactive forms of the BCKAD complex.

Determination of Branched - Chain α - Keto Acid Dehydrogenase Activity in Rat Tissues

Hyun Sook Kim,Wayne A . Johnson 생화학분자생물학회 1995 BMB Reports Vol.28 No.1

The branched-chain α-keto acid dehydrogenase (BCKAD) complex is a rate limiting enzyme which catalyzes the oxidative decarboxylation of brnnched-chain α-keto acids. Numerous studies have suggested that BCKAD is subject to covalent modification in vitro via phosphorylation and dephosphorylation, which are catalyzed by a specific kinase and phosphatase, respectively. The biggest difficulty in the assay of BCKAD activity is to arrest the interconversion between the active and inactive forms. BCKAD activity was determined from fresh rat heart and liver tissues using homogenizing and assay buffers containing inhibitors of phosphatase and kinase. The results suggest that a radiochemical assay using α-keto[1-^(14)C]-isovalerate as a substrate for the enzyme can be applied as a reliable method to determine in vitro enzyme activity with arrested interconversion between the active and inactive forms of the BCKAD complex.

Trichloroethylene induces dopaminergic neurodegeneration in Fisher 344 rats

Liu, Mei,Choi, Dong-Young,Hunter, Randy L.,Pandya, Jignesh D.,Cass, Wayne A.,Sullivan, Patrick G.,Kim, Hyoung-Chun,Gash, Don M.,Bing, Guoying Blackwell Publishing Ltd 2010 Journal of Neurochemistry Vol.112 No.3

<P><I>J. Neurochem.</I> (2010) <B>112</B>, 773–783.</P><P>Abstract</P><P>Trichloroethylene, a chlorinated solvent widely used as a degreasing agent, is a common environmental contaminant. Emerging evidence suggests that chronic exposure to trichloroethylene may contribute to the development of Parkinson’s disease. The purpose of this study was to determine if selective loss of nigrostriatal dopaminergic neurons could be reproduced by systemic exposure of adult Fisher 344 rats to trichloroethylene. In our experiments, oral administration of trichloroethylene induced a significant loss of dopaminergic neurons in the substantia nigra pars compacta in a dose-dependent manner, whereas the number of both cholinergic and GABAergic neurons were not decreased in the striatum. There was a robust decline in striatal levels of 3, 4-dihydroxyphenylacetic acid without a significant depletion of striatal dopamine. Rats treated with trichloroethylene showed defects in rotarod behavior test. We also found a significantly reduced mitochondrial complex I activity with elevated oxidative stress markers and activated microglia in the nigral area. In addition, we observed intracellular &agr;-synuclein accumulation in the dorsal motor nucleus of the vagus nerve, with some in nigral neurons, but little in neurons of cerebral cortex. Overall, our animal model exhibits some important features of Parkinsonism, and further supports that trichloroethylene may be an environmental risk factors for Parkinson’s disease.</P>

Solution-processed germanium nanowire-positioned Schottky solar cells

Yun, Ju-Hyung,Park, Yun Chang,Kim, Joondong,Lee, Hak-Joo,Anderson, Wayne A,Park, Jeunghee Springer 2011 NANOSCALE RESEARCH LETTERS Vol.6 No.1

<P>Germanium nanowire (GeNW)-positioned Schottky solar cell was fabricated by a solution process. A GeNW-containing solution was spread out onto asymmetric metal electrodes to produce a rectifying current flow. Under one-sun illumination, the GeNW-positioned Schottky solar cell yields an open-circuit voltage of 177 mV and a short-circuit current of 19.2 nA. Schottky and ohmic contacts between a single GeNW and different metal electrodes were systematically investigated. This solution process may provide a route to the cost-effective nanostructure solar architecture.</P>

Hot Populations in M87 Globular Clusters

Sohn, Sangmo T.,O'Connell, Robert W.,Kundu, Arunav,Landsman, Wayne B.,Burstein, David,Bohlin, Ralph C.,Frogel, Jay A.,Rose, James A. American Institute of Physics 2006 The Astronomical journal Vol.131 No.2

<P>To explore the production of UV-bright stars in old, metal-rich populations like those in elliptical galaxies, we have obtained Hubble Space Telescope (HST) Space Telescope Imaging Spectrograph far- and near-UV photometry of globular clusters (GCs) in four fields in the giant elliptical (gE) galaxy M87. To a limit of m<SUB>FUV</SUB> ~ 25 we detect a total of 66 GCs in common with the deep HST optical-band study of Kundu et al. Despite strong overlap in V- and I-band properties, the M87 GCs have UV-optical properties that are distinct from clusters in the Milky Way and in M31. M87 clusters, especially metal-poor ones, produce larger hot horizontal-branch populations than do Milky Way analogs. In color plots including the near-UV band, the M87 clusters appear to represent an extension of the Milky Way sequence. Cluster mass is probably not a factor in these distinctions. The most metal-rich M87 GCs in our sample are near solar metallicity and overlap the local E galaxy sample in estimated Mg<SUB>2</SUB> line indices. Nonetheless, the clusters produce much more UV light at a given Mg<SUB>2</SUB>, being up to 1 mag bluer than any gE galaxy in (FUV - V) color. The M87 GCs do not appear to represent a transition between Milky Way-type clusters and E galaxies. The differences are in the correct sense if the clusters are significantly older than the E galaxies.</P><P>Comparisons with Galactic open clusters indicate that the hot stars lie on the extreme horizontal branch, rather than being blue stragglers, and that the extreme horizontal branch becomes well populated for ages ≳5 Gyr. Existing model grids for clusters do not match the observations well, due to poorly understood giant branch mass loss or perhaps high helium abundances. We find that 41 of our UV detections have no optical-band counterparts. Most appear to be UV-bright background galaxies seen through M87. Eleven near-UV variable sources detected at only one epoch in the central field are probably classical novae. Two recurrent variable sources have no obvious explanation but could be related to activity in the relativistic jet.</P>