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Amber suppression

Stanley Ching-Cheng Huang, Amber M Smith, Bart Everts, Marco Colonna, Erika L Pearce, Joel D Schilling, Edward J Pearce
Macrophage activation status is intrinsically linked to metabolic remodeling. Macrophages stimulated by interleukin 4 (IL-4) to become alternatively (or, M2) activated increase fatty acid oxidation and oxidative phosphorylation; these metabolic changes are critical for M2 activation. Enhanced glucose utilization is also characteristic of the M2 metabolic signature. Here, we found that increased glucose utilization is essential for M2 activation. Increased glucose metabolism in IL-4-stimulated macrophages required the activation of the mTORC2 pathway, and loss of mTORC2 in macrophages suppressed tumor growth and decreased immunity to a parasitic nematode...
October 18, 2016: Immunity
Yu-Hong Cheng, Yue Zhang, Siu-Leung Chau, Samuel Kin-Man Lai, Ho-Wai Tang, Kwan-Ming Ng
Metal alloy nanoparticles (NPs) offer a new combination of unique physicochemical properties based on their pure counterparts, which can facilitate the development of novel analytical methods. Here, we demonstrated that Ag-Au alloy NPs could be utilized for optical and mass spectrometric imaging of latent fingerprints (LFPs) with improved image contrast, stability, and detection sensitivity. Upon deposition of Ag-Au alloy NPs (Ag:Au = 60:40% wt), ridge regions of the LFP became amber-coloured, while the groove regions appeared purple-blue...
October 17, 2016: ACS Applied Materials & Interfaces
Xin Li, Lung Wa Chung, Guohui Li
Fluorescent RNA aptamer Spinach can bind and activate a green fluorescent protein (GFP)-like chromophore (an anionic DFHBDI chromophore) displaying green fluorescence. Spectroscopic properties, spectral tuning, and the photoisomerization mechanism in the Spinach-DFHBDI complex have been investigated by high-level QM and hybrid ONIOM(QM:AMBER) methods (QM method: (TD)DFT, SF-BHHLYP, SAC-CI, LT-DF-LCC2, CASSCF, or MS-CASPT2), as well as classical molecular dynamics (MD) simulations. First, our benchmark calculations have shown that TD-DFT and spin-flip (SF) TD-DFT (SF-BHHLYP) failed to give a satisfactory description of absorption and emission of the anionic DFHBDI chromophore...
October 14, 2016: Journal of Chemical Theory and Computation
Roghaye Arezumand, Reza Mahdian, Sirous Zeinali, Gholamreza Hassanzadeh-Ghassabeh, Kamran Mansouri, Hossein Khanahmad, Nabiollah Namvar-Asl, Hamzeh Rahimi, Mahdi Behdani, Reza Ahangari Cohan, Mehdi Eavazalipour, Ali Ramazani, Serge Muyldermans
Placental growth factor (PlGF), a member of vascular endothelial growth factors (VEGF) family, is considered as an important antigen associated with pathological conditions such as cancer cell growth, and metastasis. PlGF-targeting via nanobody (Nb) therefore could be beneficial to modulate these pathologies. In this work, phage-display and computational approach was employed to develop a high affinity PlGF-specific Nb. An Nb library was constructed against human recombinant PlGF (rPlGF). After panning on immobilized rPlGF the periplasmic-extract (PE) of individual colonies were screened by ELISA (PE-ELISA)...
October 2016: Molecular Immunology
Ido Laskov, Paul Abou-Nader, Oreekha Amin, Charles-Andre Philip, Marie-Claude Beauchamp, Amber Yasmeen, Walter H Gotlieb
PURPOSE: Epithelial-mesenchymal transition (EMT) is a critical process for cancer metastasis and recurrence. Metformin, an effective oral antidiabetic drug, has been associated with decreased cancer risk and mortality. In this pilot study, we started to evaluate the effect of metformin on EMT in vivo and in vitro in endometrial cancer (EC). METHODS: Endometrial cancer cell lines and freshly isolated EC tumor specimens were used to assess EMT after metformin treatment...
September 2016: International Journal of Gynecological Cancer
Amber A Bokhari, Tabari M Baker, Batsukh Dorjbal, Sana Waheed, Christopher M Zahn, Chad A Hamilton, G Larry Maxwell, Viqar Syed
Nestin, an intermediate filament protein and a stem cell marker is expressed in several tumors. Until recently, little was known about the expression levels and the role of Nestin in endometrial cancer. Compared to the immortalized endometrial epithelial cell line EM-E6/E7-TERT, endometrial cancer cell lines express high to moderate levels of Nestin. Furthermore, endometrial tumors and tumor cell lines have a cancer stem-like cell subpopulation expressing CD133. Among the cancer lines, AN3CA and KLE cells exhibited both a significantly higher number of CD133+ cells and expressed Nestin at higher levels than Ishikawa cells...
September 10, 2016: Oncotarget
Scott Walker, Fiona Foster, Amber Wood, Thomas Owens, Keith Brennan, Charles H Streuli, Andrew P Gilmore
A key hallmark of cancer cells is the loss of positional control over growth and survival. Focal adhesion kinase (FAK) is a tyrosine kinase localised at sites of integrin-mediated cell adhesion to the extracellular matrix. FAK controls a number of adhesion-dependent cellular functions, including migration, proliferation and survival. Although FAK is overexpressed and activated in metastatic tumours, where it promotes invasion, it can also be elevated in cancers that have yet to become invasive. The contribution of FAK to the early stages of tumourigenesis is not known...
September 6, 2016: Oncotarget
Andre Maranhao, Andrew D Ellington
There have been considerable advancements in the incorporation of non-canonical amino acids (ncAA) into proteins over the last two decades. The most widely used method for site-specific incorporation of non-canonical amino acids, amber stop codon suppression, typically employs an orthogonal translation system (OTS) consisting of a heterologous aminoacyl-tRNA synthetase:tRNA pair that can potentially expand an organism's genetic code. However, the orthogonal machinery sometimes imposes fitness costs on an organism, in part due to mischarging and a lack of specificity...
September 7, 2016: ACS Synthetic Biology
Yufan Wu, Steven G Boxer
The vibrational Stark effect (VSE) has been used to measure the electric field in the active site of ketosteroid isomerase (KSI). These measured fields correlate with ΔG(⧧) in a series of conventional mutants, yielding an estimate for the electrostatic contribution to catalysis (Fried et al. Science 2014, 346, 1510-1513). In this work we test this result with much more conservative variants in which individual Tyr residues in the active site are replaced by 3-chlorotyrosine via amber suppression. The electric fields sensed at the position of the carbonyl bond involved in charge displacement during catalysis were characterized using the VSE, where the field sensitivity has been calibrated by vibrational Stark spectroscopy, solvatochromism, and MD simulations...
September 14, 2016: Journal of the American Chemical Society
Lulu Mao, Whitney Summers, Shulin Xiang, Lin Yuan, Robert T Dauchy, Amberly Reynolds, Melissa A Wren-Dail, David Pointer, Tripp Frasch, David E Blask, Steven M Hill
: The importance of the circadian/melatonin signal in suppressing the metastatic progression of breast and other cancers has been reported by numerous laboratories including our own. Currently, the mechanisms underlying the anti-metastatic actions of melatonin have not been well established. In the present study, the anti-metastatic actions of melatonin were evaluated and compared on the ERa-negative, Her2-positive SKBR-3 breast tumor cell line and ERa-positive MCF-7 cells overexpressing a constitutively active HER2...
August 17, 2016: Molecular Cancer Research: MCR
Amber W Kinsey, Stacy R Cappadona, Lynn B Panton, Brittany R Allman, Robert J Contreras, Robert C Hickner, Michael J Ormsbee
We have previously shown that ingesting protein at night before sleep is either beneficial or non-detrimental to metabolism, health, and body composition in obese women. However, the overnight protein-induced lipolytic actions and mechanism for improved metabolism and body composition have not been fully established. Therefore, in a crossover design, twelve obese men (age, 27.0 ± 2.2 years) were randomly assigned to ingest (within 30 min of sleep) casein protein (CAS, 120 kcal) or a non-nutritive placebo (PLA) before going to sleep...
2016: Nutrients
Robert B Quast, Andrei Sonnabend, Marlitt Stech, Doreen A Wüstenhagen, Stefan Kubick
Cell-free protein synthesis systems derived from eukaryotic sources often provide comparatively low amounts of several μg per ml of de novo synthesized membrane protein. In order to overcome this, we herein demonstrate the high-yield cell-free synthesis of the human EGFR in a microsome-containing system derived from cultured Sf21 cells. Yields were increased more than 100-fold to more than 285 μg/ml by combination of IRES-mediated protein translation with a continuous exchange cell-free reaction format that allowed for prolonged reaction lifetimes exceeding 24 hours...
2016: Scientific Reports
Stephanie Bonney, Susan Harrison-Uy, Swati Mishra, Amber M MacPherson, Youngshik Choe, Dan Li, Shou-Ching Jaminet, Marcus Fruttiger, Samuel J Pleasure, Julie A Siegenthaler
UNLABELLED: As neural structures grow in size and increase metabolic demand, the CNS vasculature undergoes extensive growth, remodeling, and maturation. Signals from neural tissue act on endothelial cells to stimulate blood vessel ingression, vessel patterning, and acquisition of mature brain vascular traits, most notably the blood-brain barrier. Using mouse genetic and in vitro approaches, we identified retinoic acid (RA) as an important regulator of brain vascular development via non-cell-autonomous and cell-autonomous regulation of endothelial WNT signaling...
July 20, 2016: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
Russell K Monson, Amberly A Neice, Nicole A Trahan, Ian Shiach, Joel T McCorkel, David J P Moore
Plant isoprene emissions have been linked to several reaction pathways involved in atmospheric photochemistry. Evidence exists from a limited set of past observations that isoprene emission rate (Is ) decreases as a function of increasing atmospheric CO2 concentration, and that increased temperature suppresses the CO2 effect. We studied interactions between intercellular CO2 concentration (Ci ) and temperature as they affect Is in field-grown hybrid poplar trees in one of the warmest climates on earth - the Sonoran Desert of the southwestern United States...
June 28, 2016: Plant, Cell & Environment
Matthew R Gardner, Christoph H Fellinger, Neha R Prasad, Amber S Zhou, Hema R Kondur, Vinita R Joshi, Brian D Quinlan, Michael Farzan
UNLABELLED: The HIV-1 envelope glycoprotein (Env) is a trimer of gp120/gp41 heterodimers that mediates viral entry. Env binds cellular CD4, an association which stabilizes a conformation favorable to its subsequent association with a coreceptor, typically CCR5 or CXCR4. The CD4- and coreceptor-binding sites serve as epitopes for two classes of HIV-1-neutralizing antibodies: CD4-binding site (CD4bs) and CD4-induced (CD4i) antibodies, respectively. Here we observed that, at a fixed total concentration, mixtures of the CD4i antibodies (E51 or 412d) and the CD4bs antibody VRC01 neutralized the HIV-1 isolates 89...
September 1, 2016: Journal of Virology
Atsushi Ogawa, Junichiro Tabuchi, Yasunori Doi, Masashi Takamatsu
We have developed a novel type of biofunction-assisted, signal-turn-on sensor for simply and homogenously detecting DNA. This sensor system is composed of two types of in vitro-transcribed label-free RNAs (a 3' premature amber suppressor tRNA probe and an amber-mutated mRNA encoding a reporter protein), RNase H, and a wheat germ extract (WGE). A target DNA induces the 3' end maturation of the tRNA probe, which is enhanced by RNase H and leads to the expression of a full-length reporter protein through amber suppression in WGE, while there is almost no expression without the target due to the inactivity of the premature probe...
August 1, 2016: Bioorganic & Medicinal Chemistry Letters
Lauren N Guthrie, Kavitha Abiraman, Emily S Plyler, Neil T Sprenkle, Sara A Gibson, Braden C McFarland, Rajani Rajbhandari, Amber L Rowse, Etty N Benveniste, Gordon P Meares
Inflammation and endoplasmic reticulum (ER) stress are associated with many neurological diseases. ER stress is brought on by the accumulation of misfolded proteins in the ER, which leads to activation of the unfolded protein response (UPR), a conserved pathway that transmits signals to restore homeostasis or eliminate the irreparably damaged cell. We provide evidence that inhibition or genetic haploinsufficiency of protein kinase R-like endoplasmic reticulum kinase (PERK) can selectively control inflammation brought on by ER stress without impinging on UPR-dependent survival and adaptive responses or normal immune responses...
July 22, 2016: Journal of Biological Chemistry
Xiaobao Bi, Kalyan Kumar Pasunooti, Ahmad Hussen Tareq, John Takyi-Williams, Chuan-Fa Liu
Here we report a new site-specific conjugation strategy to modify proteins via thiazolidine ligation. Proteins harbouring a 1,2-aminothiol moiety introduced by amber codon suppression technology could be modified chemoselectively with aldehyde-functionalized reagents, such as a biotin-labeled peptide or ubiquitin, under mild conditions to yield homogeneous biotinylated or ubiquitinated products.
June 21, 2016: Organic & Biomolecular Chemistry
Brittany Mathias, Amber L Delmas, Tezcan Ozrazgat-Baslanti, Erin L Vanzant, Benjamin E Szpila, Alicia M Mohr, Frederick A Moore, Scott C Brakenridge, Babette A Brumback, Lyle L Moldawer, Philip A Efron
OBJECTIVE: We hypothesized that after sepsis in humans, MDSCs will be persistently increased, functionally immunosuppressive, and associated with adverse clinical outcomes. BACKGROUND: Cancer and sepsis have surprisingly similar immunologic responses and equally dismal long term consequences. In cancer, increased myeloid-derived suppressor cells (MDSCs) induce detrimental immunosuppression, but little is known about the role of MDSCs after sepsis. METHODS: Blood was obtained from 74 patients within 12 hours of severe sepsis/septic shock (SS/SS), and at set intervals out to 28 days, and also in 18 healthy controls...
May 9, 2016: Annals of Surgery
Rachael V McCaleb, Arpita S Gandhi, Stephen Michael Clark, Amber B Clemmons
BACKGROUND: Acid suppressive therapy (AST)-namely, proton pump inhibitors (PPIs) and histamine-2 receptor antagonists (H2RAs)-is routinely prescribed to hospitalized patients for stress ulcer prophylaxis (SUP). OBJECTIVE: To identify the incidence of and indications for AST use in the hematology/oncology population as well as to identify the occurrence of the following PPI-associated adverse events: pneumonia and Clostridium difficile-associated diarrhea (CDAD)...
July 2016: Annals of Pharmacotherapy
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