Biochemistry, Biophysics, and Structural Biology Commons^™

Mcnamara 201412 Nih Scap Innocentive Challenge Solution - T-Bow Rainbow T-Cells And Tumor Cells Spatial Multiplexing Gene Expression Reporter System – Plus Supplement Plus Posters - 20151027 - Please Download "75" Instead, George Mcnamara

George McNamara

McNamara 201412 NIH SCAP InnoCentive Challenge Solution - T-Bow Rainbow T-cells and Tumor Cells Spatial Multiplexing Gene Expression Reporter System – plus supplement plus posters - 20151027.

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Please download the current 20151027 (October 27, 2015) Tattletales and T-Bow update from

http://works.bepress.com/gmcnamara/75/

The bepress web site is not letting me replace the old pdf here at "65" with the additional 10 pages update.

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The download is my/Cooper lab solution (submission) to the 2014 NIH Single Cell Analysis Program (SCAP) InnoCentive Challenge, "Follow That Cell". I submitted the Solution on 20141215Mon (with 20 minutes to spare). The Challenge web page …

Tattletales And T-Bow Update 20151027tue, George Mcnamara

George McNamara

20151027Tue this "75"

http://works.bepress.com/gmcnamara/75

is my update of "65" posting

See text at

http://works.bepress.com/gmcnamara/65/

for text summary. The PDf here in "75" supersedes "65".

The PDF here has 10 pages added to the end from the "65" version (pages 40-49 of PDF when including the bepress cover page)..

here is the text in my cover page (bepress may add its own cover):

20151027Tue: added 10 page e-poster at bottom explaining Binary Tattletales and T-Bow. That is, binary with respect to protein components. For one color (number of repeats, epitope tags, FPs are examples, here rounded to convenient numbers):

1. 100 …

Experimental Demonstration Of Bindingless Signal Delivery In Human Cells Via Microfluidics, Fang-Tzu Chuang

Fang-Tzu Chuang

The cellular signal transduction is commonly believed to rely on the direct “contact” or “binding” of the participating molecule reaction that depends positively on the corresponding molecule concentrations. In living systems, however, it is somewhat difficult to precisely match the corresponding rapid “binding,” depending on the probability of molecular collision, existing in the cellular receptor-ligand interactions. Thus, a question arises that if there is another mechanism (i.e., bindingless) that could promote this signal communication. According to this hypothesis, we report a cellular model based on the examination of intracellular calcium concentration to explore whether the unidentified signal delivery in cells …

Stellaris Fishing 20131125mon Part 2 Of 2, George Mcnamara

George McNamara

Stellaris FISH dataset using three FISH probe sets.

Slides courtesy of Biosearch Technologies,

https://www.biosearchtech.com/store/product.aspx?catid=224,318,324

see http://stellarisfish.smugmug.com/ for online gallery by Biosearch.

This experiment was to evaluate the crosstalk between the Biosearch fluorophores:

Quasar 570

CAL Fluor Red 610 (CFR 610)

Quasar 670

DAPI (DNA counterstain)

Autofluorescence (green, but sometimes showing up in other channels).

and our lab's Leica DMI6000 fluorescence microscope with Leica filter sets:

DAPI

GFP (L5)

Cy3 (N3)

Texas Red (TxRed2)

Cy5 (Y5)

I also acquired green channel and red channel with exciter filters in our ASI excitation wheel:

GFP + 492 exciter

Texas Red (TxRed2) + 572 …

Enhanced Transmission Of Higher Order Plasmon Modes With Random Au Nanoparticles In Periodic Hole Arrays, Fang-Tzu Chuang

Fang-Tzu Chuang

The enhanced transmission of higher order plasmon modes with random gold nanoparticles embedded in periodic hole arrays using asymmetric pair aperture as a unit is investigated in the midinfrared regime. Different thicknesses of gold film were deposited inside holes and then annealed to form randomly sized and distributed nanoparticles. The holes deposited with thin gold film exhibit significantly enhanced transmission in higher order modes after thermal annealing. The enormous local electric field around the nanoparticles enhances the scattering effect that contributes to the enhanced infrared transmission. This unique design, which integrates localized and propagating surface plasmons, provides an easy way …

Microfabricated Nanotopological Surfaces For Study Of Adhesion-Dependent Cell Mechanosensitivity, Weiqiang Chen, Yubing Sun, Jianping Fu

Weiqiang Chen

Cells exhibit high sensitivity and diverse responses to the intrinsic nanotopography of the extracellular matrix through their nanoscale cellular sensing machinery. A simple microfabrication method for precise control and spatial patterning of the local nanoroughness on glass surfaces by using photolithography and reactive ion etching is reported. It is demonstrated that local nanoroughness as a biophysical cue could regulate a diverse array of NIH/3T3 fi broblast behaviors, including cell morphology, adhesion, proliferation, migration, and cytoskeleton contractility. The capability to control and further predict cellular responses to nanoroughness might suggest novel methods for developing biomaterials mimicking nanotopographic structures in vivo for …

Bone Morphogenetic Protein 4 Inhibition Of Tgfβ-2 Induced Cross-Linked Actin Networks In Human Trabecular Meshwork Cells, Rafael A. Lima, Michela Montecchi-Palmer, Jaclyn Bermudez, Abbot F. Clark Phd, Robert J. Wordinger Phd

ral5@rice.edu

Bone Morphogenetic Protein 4 Inhibition of TGFβ-2 Induced Cross-Linked Actin Networks (CLANs) in Human Trabecular Meshwork Cells

RAFAEL A. LIMA1, MICHELA MONTECCHI-PALMER2,3, JACLYN BERMUDEZ2 ABBOT F. CLARK2, AND ROBERT J. WORDINGER2

1 Rice University, Houston, Texas, 2 Department of Cell Biology and Anatomy, North Texas Eye Research Institute, University of North Texas Health Science Center, Fort Worth, Texas, 3 Alcon Research, Ltd., Fort Worth, Texas

Background:

Primary open-angle glaucoma (POAG) is a degenerative eye disease characterized by irreversible loss of retinal ganglion cells. One of the most significant risk factors for POAG is increased intraocular pressure (IOP). Elevated IOP is …

Nanoroughened Surfaces For Efficient Capture Of Circulating Tumor Cells Without Using Capture Antibodies, Weiqiang Chen, Shinuo Weng, Feng Zhang, Steven Allen, Xiang Li, Liwei Bao, Raymond H. W. Lam, Jill A. Macoska, Sofia D. Merajver, Jianping Fu

Weiqiang Chen

Circulating tumor cells (CTCs) detached from both primary and metastatic lesions represent a potential alternative to invasive biopsies as a source of tumor tissue for the detection, characterization and monitoring of cancers. Here we report a simple yet effective strategy for capturing CTCs without using capture antibodies. Our method uniquely utilized the differential adhesion preference of cancer cells to nanorough surfaces when compared to normal blood cells and thus did not depend on their physical size or surface protein expression, a significant advantage as compared to other existing CTC capture techniques.

Effect Of Paired Apertures In A Periodic Hole Array On Higher Order Plasmon Modes, Fang-Tzu Chuang

Fang-Tzu Chuang

We demonstrate that the transmission of higherorder surface plasmon modes in the mid-infrared range can be enhanced through rectangular hole array on the basis of paired apertures. Experiments prove that enhanced high-order transmission can be generated by either identical shapes or combinations of different hole shapes in pairs. The structure factor is adopted to explain the observed intensity of enhanced transmission. Numerical simulations of the enhanced secondorder mode verify a significant field enhancement in a unit cell of pairs. It is clarified that the separation between the paired apertures and the paired resonance is the key to determine certain higher-order …

Two Infrared Emission Modes With Different Wavelengths And Orthogonal Polarization In A Waveguide Thermal Emitter, Fang-Tzu Chuang

Fang-Tzu Chuang

This study investigates a Au/SiO2/Au waveguide thermal emitter incorporating a metallic grating embedded in the SiO2 layer. The metal grating acts as a beam splitter, dividing the device into two waveguide structures determined by the polarization of the waveguide modes. The thermal radiation spectrum exhibits two peaks with orthogonal polarization. The emitted wavelengths of the two waveguide modes can be adjusted by controlling the thickness of the SiO2 layers on both sides of the metallic grating during the fabrication process. The emission peaks have ratios of the full width at half maximum to the peak wavelength of 0.057 and 0.05 …

Libraries At The University Of Massachusetts Amherst: Seeking An International Perspective, Maxine G. Schmidt

Maxine G Schmidt

Presentation delivered to librarians in China, Japan and South Korea as part of my sabbatical research on the use of libraries by Asian students in their home countries.

Nanoprojection Lithography Using Self-Assembled Interference Modules For Manufacturing Plasmonic Gratings, Fang-Tzu Chuang

Fang-Tzu Chuang

Abstract—A new nanoprojection lithography (NPL) is proposed to manufacture plasmonic nanogratings. Here, low-cost self-assembly elastomeric polydimethylsiloxane wavy structures are used as the interference modules, with their periods and amplitudes largely controlled by the applied mechanical strain in the synthesis process. Well defined plasmonic grating couplers with desired feature sizes and wavelengths of operation were obtained. This NPL may enable large-area and flash manufacturing of plasmonic nanogratings with tunable array periods.

Nanotopography Influences Adhesion, Spreading, And Self-Renewal Of Human Embryonic Stem Cells, Weiqiang Chen, Luis G. Villa-Diaz, Yubing Sun, Shinuo Weng, Jin Koo Kim, Raymond H. W. Lam, Lin Han, Rong Fan, Paul H. Krebsbach, Jianping Fu

Weiqiang Chen

Human embryonic stem cells (hESCs) have great potentials for future cell-based therapeutics. However, their mechanosensitivity to biophysical signals from the cellular microenvironment is not well characterized. Here we introduced an effective microfabrication strategy for accurate control and patterning of nanoroughness on glass surfaces. Our results demonstrated that nanotopography could provide a potent regulatory signal over different hESC behaviors, including cell morphology, adhesion, proliferation, clonal expansion, and self-renewal. Our results indicated that topological sensing of hESCs might include feedback regulation involving mechanosensory integrin-mediated cell matrix adhesion, myosin II, and E-cadherin. Our results also demonstrated that cellular responses to nanotopography were cell-type …

Integrative Bayesian Analysis Of High-Dimensional Multi-Platform Genomics Data, Wenting Wang, Veerabhadran Baladandayuthapani, Jeffrey S. Morris, Bradley M. Broom, Ganiraju C. Manyam, Kim-Anh Do

Jeffrey S. Morris

Motivation: Analyzing data from multi-platform genomics experiments combined with patients’ clinical outcomes helps us understand the complex biological processes that characterize a disease, as well as how these processes relate to the development of the disease. Current integration approaches that treat the data are limited in that they do not consider the fundamental biological relationships that exist among the data from platforms.

Statistical Model: We propose an integrative Bayesian analysis of genomics data (iBAG) framework for identifying important genes/biomarkers that are associated with clinical outcome. This framework uses a hierarchical modeling technique to combine the data obtained from multiple platforms …

Hslic Fall 2012 Scholarship Committee Report, Ann Jordan

Ann Jordan

No abstract provided.

Arcane Secrets Of The Umass Libraries, Maxine G. Schmidt

Maxine G Schmidt

No abstract provided.

Self-Assembled Rippling Structure Based On Metal-Elastomer Composite For Tunable Plasmonics, Fang-Tzu Chuang

Fang-Tzu Chuang

We demonstrate that a periodic metallic wavy-structure can be easily fabricated on elastomeric polydymethylsiloxane (PDMS) substrates. The metallic wavy-structures are self-assembly formed by thermal evaporation of Ag on PDMS without any lithography processes. The size of the PDMS substrate with an aspect ratio (the ratio of length to width) higher than 2 are found to be necessary to self-assemble the one-dimensional metallic wavy-structure. The period of the wavy-structure is controllable by varing the thickness of the deposited silver film. The elastomeric- gratings can excite surface plasmon on Ag/air interfaces; the resonant wavelength can be tuned to a range of 3.5% …

Science Boot Camp For Librarians: Cpd On A Shoestring, Maxine G. Schmidt, Rebecca Reznik-Zellen

Maxine G Schmidt

Science Boot Camp for Librarians was envisioned as a casual but intensive immersion event into selected scientific subjects that employ networked computing capabilities for research and collaboration. The goal of the event is to provide librarians with networking opportunities, but more importantly, to give them some of the context and ocabulary of a discipline to enable them to better engage faculty and research scientists with regard to escience. A half-day is devoted to each of three topics chosen for that year’s camp. A local faculty member provides an overview of the research area, and a second describes a single project …

Localized Shape Resonance On Silver Film Perforated By H-Shaped And More Complex Shaped Hole Arrays, Fang-Tzu Chuang

Fang-Tzu Chuang

The experimental results of light transmission through periodic array of H-shaped hole and more complicated hole which is a combination of multiple U shape are demonstrated. The observations indicate that the localized shape resonance in the longest resonant length of unfolded U-shaped part of the hole always appears. However, localized modes resonant in smaller U-shaped length don't always appear. Localized mode with non-U-shaped resonant path cannot be seen in our sample. In addition, localized mode with different order and resonant path can be excited by different polarized light.

Ppar Agonists Down-Regulate The Expression Of Atp10c Mrna During Adipogenesis, A Peretich, Maria Cekanova Ms, Rndr, Phd, S Hurst, Sj Baek, Madhu Dahr

Maria Cekanova MS, RNDr, PhD

No abstract provided.

E-Science @ Umass: Anticipating And Supporting E-Science Activities At The University Of Massachusetts, Maxine G. Schmidt, Rebecca Reznik-Zellen

Maxine G Schmidt

In March of 2008 an Ad Hoccommittee of Science Librarians from the University of Massachusetts Five Campus System convened to discuss the challenges of e-science and prepare the Libraries for their role in e-science initiatives. Three primary outcomes intended to support e-science activities emerged from the work of the Ad Hoc committee.

E-Science @ The University Of Massachusetts, Maxine G. Schmidt, Rebecca Reznik-Zellen, Raquel Rivera, Cecilia P. Mullen

Maxine G Schmidt

e-Science @ the University of Massachusetts Abstract: What is e-Science and how can libraries and librarians support it? The University of Massachusetts takes a proactive approach to support network-enabled research on its campuses and provides examples where e-Science is already at work. Statement: “e-Science” is a term commonly used to describe research in a networked environment, a growing trend not only in the sciences, but the arts and humanities as well. e-Science creates both opportunities and challenges for academic libraries. The opportunities lie in leveraging the basic skill set that libraries and librarians already possess: the knowledge of and practical …

Umass Libraries 2009, Maxine G. Schmidt

Maxine G Schmidt

No abstract provided.

Important, But Odd And Obscure, Reasons To Use The Library, Maxine G. Schmidt

Maxine G Schmidt

No abstract provided.

Expression Of G-Protein Inwardly Rectifying Potassium Channels (Girks) In Lung Cancer Cell Lines, Howard Plummer 3rd, Madhu Dhar, Maria Cekanova Ms, Rndr, Phd, Hildegard Schuller

Maria Cekanova MS, RNDr, PhD

BACKGROUND: Previous data from our laboratory has indicated that there is a functional link between the beta-adrenergic receptor signaling pathway and the G-protein inwardly rectifying potassium channel (GIRK1) in human breast cancer cell lines. We wanted to determine if GIRK channels were expressed in lung cancers and if a similar link exists in lung cancer. METHODS: GIRK1-4 expression and levels were determined by reverse transcription polymerase chain reaction (RT-PCR) and real-time PCR. GIRK protein levels were determined by western blots and cell proliferation was determined by a 5-bromo-2'-deoxyuridine (BrdU) assay. RESULTS: GIRK1 mRNA was expressed in three of six small …

Molecular Characterization Of A Isoenzyme Of The Targeting Peptide Degrading Protease, Prep2- Catalysis, Subcellular Localization, Expression And Evolution, S. Bhushan, A. Stahl, S. Nilsson, B. Lefebvre, D. Mcwilliams, S.J. Wright, M. Seki, D.A. Liberles, K. Shinozaki, Barry D. Bruce, M. Boutry, E. Glaser

Barry D. Bruce

We have previously identified a zinc metalloprotease involved in the degradation of mitochondrial and chloroplast targeting peptides, the presequence protease (PreP). In the Arabidopsis thaliana genomic database, there are two genes that correspond to the protease, the zinc metalloprotease (AAL90904) and the putative zinc metalloprotease (AAG13049). We have named the corresponding proteins AtPreP1 and AtPreP2, respectively. AtPreP1 and AtPreP2 show significant differences in their targeting peptides and the proteins are predicted to be localized in different compartments. AtPreP1 was shown to degrade both mitochondrial and chloroplast targeting peptides and to be dual targeted to both organelles using an ambiguous targeting …

Multiple Luteinizing Hormone Receptor (Lhr) Protein Variants, Interspecies Reactivity Of Anti-Lhr Mab Clone 3b5, Subcellular Localization Of Lhr In Human Placenta, Pelvic Floor And Brain, And Possible Role For Lhr In The Development Of Abnormal Pregnancy, Pelvic Floor Disorders And Alzheimer's Disease, A Bukovsky, K Indrapichate, H Fujiwara, Maria Cekanova Ms, Rndr, Phd, Me Ayala, R Dominguez, Mr Caudle, J Wimalsena, Rf Elder, P Copas, Jf Foster, Ri Fernando, Dc Henley, Nb Upadhyaya

Maria Cekanova MS, RNDr, PhD

Distinct luteinizing hormone receptor (LHR) protein variants exist due to the posttranslational modifications. Besides ovaries, LHR immunoreactivity (LHRI) was also found in other tissues, such as the brain, fallopian tube, endometrium, trophoblast and resident tissue macrophages. The 3B5 mouse monoclonal antibody was raised against purified rat LHR. In rat, porcine and human ovaries, the 3B5 identified six distinct LHR bands migrating at approximately 92, 80, 68, 59, 52 and 48 kDa. Characteristic LHRI was detected in rat, human and porcine corpora lutea. During cellular differentiation, subcellular LHR distribution changed from none to granular cytoplasmic, perinuclear, surface, nuclear and no staining. …

Expression And Localization Of Estrogen Receptor-Alpha Protein In Normal And Abnormal Term Placentae And Stimulation Of Trophoblast Differentiation By Estradiol, A Bukovsky, Maria Cekanova Ms, Rndr, Phd, Mr Caudle, J Wimalasena, Js Foster, Dc Henley, Rf Elder

Maria Cekanova MS, RNDr, PhD

Estrogens play an important role in the regulation of placental function, and 17-beta-estradiol (E2) production rises eighty fold during human pregnancy. Although term placenta has been found to specifically bind estrogens, cellular localization of estrogen receptor alpha (ER-alpha) in trophoblast remains unclear. We used western blot analysis and immunohistochemistry with h-151 and ID5 monoclonal antibodies to determine the expression and cellular localization of ER-alpha protein in human placentae and cultured trophoblast cells. Western blot analysis revealed a ~65 kDa ER-alpha band in MCF-7 breast carcinoma cells (positive control). A similar band was detected in five normal term placentae exhibiting strong …

Placental Expression Of Estrogen Receptor Beta And Its Hormone Binding Variant – Comparison With Estrogen Receptor Alpha And A Role For Estrogen Receptors In Asymmetric Division And Differentiation Of Estrogen-Dependent Cells, Antonin Bukovsky, Michael R. Caudle, Maria Cekanova Ms, Rndr, Phd, Romaine I. Fernando, Jay Wimalasena, James S. Foster, Donald C. Henley, Robert F. Elder

Maria Cekanova MS, RNDr, PhD

During human pregnancy, the production of 17-beta-estradiol (E2) rises steadily to eighty fold at term, and placenta has been found to specifically bind estrogens. We have recently demonstrated the expression of estrogen receptor alpha (ER-alpha) protein in human placenta and its localization in villous cytotrophoblast (CT), vascular pericytes, and amniotic fibroblasts. In vitro, E2 stimulated development of large syncytiotrophoblast (ST) aggregates. In the present study we utilized ER-beta affinity purified polyclonal (N19:sc6820) and ER-alpha monoclonal (clone h-151) antibodies. Western blot analysis revealed a single ~52 kDa ER-beta band in chorionic villi (CV) protein extracts. In CV, strong cytoplasmic ER-beta immunoreactivity …

Variability Of Placental Expression Of Cyclin E Low Molecular Weight Variants, A Bukovsky, Maria Cekanova Ms, Rndr, Phd, Mr Caudle, J Wimalasena, Js Foster, Ja Keenan, Rf Elder

Maria Cekanova MS, RNDr, PhD

No abstract provided.