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https://www.readbyqxmd.com/read/28723211/improving-fore-a-new-inlet-design-for-filtering-samples-through-individual-microarray-spots
#1
Victoria de Lange, Marco Habegger, Marco Schmidt, János Vörös
In this publication we present an improvement to our previously introduced vertical flow microarray, the FoRe array, which capitalizes on the fusion of immunofiltration and densely packed micron test sites. Filtering samples through individual microarray spots allows us to rapidly analyze dilute samples with high-throughput and high signal-to-noise. Unlike other flowthrough microarrays, in the FoRe design samples are injected into micron channels and sequentially exposed to different targets. This arrangement makes it possible to increase the sensitivity of the microarray by simply increasing the sample volume or to rapidly reconcentrate samples after preprocessing steps dilute the analyte...
March 24, 2017: ACS Sensors
https://www.readbyqxmd.com/read/28723140/rapid-and-label-free-detection-of-interferon-gamma-via-an-electrochemical-aptasensor-comprising-a-ternary-surface-monolayer-on-a-gold-interdigitated-electrode-array
#2
Shaowei Ding, Curtis Mosher, Xian Y Lee, Suprem R Das, Allison A Cargill, Xiaohui Tang, Bolin Chen, Eric S McLamore, Carmen Gomes, Jesse M Hostetter, Jonathan C Claussen
A label-free electrochemical impedance spectroscopy (EIS) aptasensor for rapid detection (<35 min) of interferon-gamma (IFN-γ) was fabricated by immobilizing a RNA aptamer capture probe (ACP), selective to IFN-γ, on a gold interdigitated electrode array (Au IDE). The ACP was modified with a thiol group at the 5' terminal end and subsequently co-immobilized with 1,6-hexanedithiol (HDT) and 6-mercapto-1-hexanolphosphate (MCH) to the gold surface through thiol-gold interactions. This ACP/HDT-MCH ternary surface monolayer facilitates efficient hybridization with IFN-γ and displays high resistance to nonspecific adsorption of nontarget proteins [i...
February 24, 2017: ACS Sensors
https://www.readbyqxmd.com/read/28722749/rho-associated-protein-kinases-as-therapeutic-targets-for-both-vascular-and-parenchymal-pathologies-in-alzheimer-s-disease
#3
REVIEW
Aaron Y Lai, JoAnne McLaurin
The causes of late-onset Alzheimer's disease are unclear and likely multifactorial. Rho-associated protein kinases (ROCKs) are ubiquitously expressed signaling messengers that mediate a wide array of cellular processes. Interestingly, they play an important role in several vascular and brain pathologies implicated in Alzheimer's etiology, including hypertension, hypercholesterolemia, blood-brain barrier disruption, oxidative stress, deposition of vascular and parenchymal amyloid-beta peptides, tau hyperphosphorylation, and cognitive decline...
July 19, 2017: Journal of Neurochemistry
https://www.readbyqxmd.com/read/28722183/a-rare-polyglycine-type-ii-like-helix-motif-in-naturally-occurring-proteins
#4
Eberhard Warkentin, Sina Weidenweber, Karola Schühle, Ulrike Demmer, Johann Heider, Ulrich Ermler
Common structural elements in proteins such as α-helices or β-sheets are characterized by uniformly repeating, energetically favorable main chain conformations which additionally exhibit a completely saturated hydrogen-bonding network of the main chain NH and CO groups. Although polyproline or polyglycine type II helices (PPII or PGII ) are frequently found in proteins, they are not considered as equivalent secondary structure elements because they do not form a similar self-contained hydrogen-bonding network of the main chain atoms...
July 19, 2017: Proteins
https://www.readbyqxmd.com/read/28722147/surface-isoelectric-focusing-sief-with-carrier-ampholyte-ph-gradient
#5
Zhichao Wang, Cornelius Ivory, Adrienne R Minerick
Isoelectric focusing (IEF) is a powerful tool for amphoteric protein separations because of high sensitivity, bio-compatibility, and reduced complexity compared to chromatography or mechanical separation techniques. IEF miniaturization is attractive because it enables rapid analysis, easier adaptation to point of care applications, and smaller sample demands. However, existing small-scale IEF tools have not yet been able to analyze single protein spots from array libraries, which are ubiquitous in many pharmaceutical discovery and screening protocols...
July 19, 2017: Electrophoresis
https://www.readbyqxmd.com/read/28720665/targeted-exome-sequencing-of-krebs-cycle-genes-reveals-candidate-cancer-predisposing-mutations-in-pheochromocytomas-and-paragangliomas
#6
Laura Remacha, Iñaki Comino-Méndez, Susan Richter, Laura Contreras, Maria Currás-Freixes, Guillermo Pita, Rocío Letón, Antonio Galarreta, Rafael Torres-Pérez, Emiliano Honrado, Scherezade Jiménez, Lorena Maestre, Sebastian Moran, Manel Esteller, Jorgina Satrústegui, Graeme Eisenhofer, Mercedes Robledo, Alberto Cascon
Purpose: Mutations in Krebs cycle genes are frequently found in patients with pheochromocytomas/paragangliomas. Disruption of SDH, FH or MDH2 enzymatic activities lead to accumulation of specific metabolites, which give rise to epigenetic changes in the genome that cause a characteristic hypermethylated phenotype. Tumors showing this phenotype, but no alterations in the known predisposing genes, could harbor mutations in other Krebs cycle genes. <p>Experimental Design: We used downregulation and methylation of RBP1, as a marker of a hypermethylation phenotype, to select eleven pheochromocytomas and paragangliomas for targeted exome sequencing of a panel of Krebs cycle-related genes...
July 18, 2017: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/28720649/the-greatest-kinetochore-show-on-earth
#7
Gerben Vader, Andrea Musacchio
Coordinated chromosome duplication and segregation is key to the existence of every organism on our planet. In eukaryotes, sophisticated protein assemblies called kinetochores are universally required for chromosome segregation, but their protein composition can diverge across the eukaryotic tree of life. In this issue of EMBO Reports, van Hooff et al [1] shed light on kinetochore evolution with a comprehensive study of kinetochore composition across 90 phylogenetically diverse eukaryotes. They show that certain kinetochore complexes have taken distinct evolutionary paths to arrive at a strikingly broad compositional array in present-day eukaryotes, providing exciting new insights into the origins, function, and flexibility of eukaryotic kinetochores...
July 18, 2017: EMBO Reports
https://www.readbyqxmd.com/read/28719984/cytotoxic-and-immunomodulatory-potential-activity-of-physalis-peruviana-fruit-extracts-on-cervical-cancer-hela-and-fibroblast-l929-cells
#8
Helen Mier-Giraldo, Luis Eduardo Díaz-Barrera, Lucy Gabriela Delgado-Murcia, Manuel Fernando Valero-Valdivieso, Gabriela Cáez-Ramírez
It was purposed to evaluate the biological potential of ethanol and isopropanol crude extracts of ripe Physalis peruviana fruits. Cytotoxic and immunomodulatory effects of the expression of interleukin-6, interleukin-8, and monocyte chemoattractant protein-1 (MCP-1) were evaluated on human cervical cancer (HeLa) and murine fibroblast (L929) cells. The composition was evaluated by high-performance liquid chromatography diode-array detection and high-performance liquid chromatography ultraviolet/visible detection...
January 1, 2017: Journal of Evidence-based Complementary & Alternative Medicine
https://www.readbyqxmd.com/read/28719003/mutation-intolerant-genes-and-targets-of-fmrp-are-enriched-for-nonsynonymous-alleles-in-schizophrenia
#9
Ganna Leonenko, Alexander L Richards, James T Walters, Andrew Pocklington, Kimberly Chambert, Mariam M Al Eissa, Sally I Sharp, Niamh L O'Brien, David Curtis, Nicholas J Bass, Andrew McQuillin, Christina Hultman, Jennifer L Moran, Steven A McCarroll, Pamela Sklar, Benjamin M Neale, Peter A Holmans, Michael J Owen, Patrick F Sullivan, Michael C O'Donovan
Risk of schizophrenia is conferred by alleles occurring across the full spectrum of frequencies from common SNPs of weak effect through to ultra rare alleles, some of which may be moderately to highly penetrant. Previous studies have suggested that some of the risk of schizophrenia is attributable to uncommon alleles represented on Illumina exome arrays. Here, we present the largest study of exomic variation in schizophrenia to date, using samples from the United Kingdom and Sweden (10,011 schizophrenia cases and 13,791 controls)...
July 18, 2017: American Journal of Medical Genetics. Part B, Neuropsychiatric Genetics
https://www.readbyqxmd.com/read/28718891/mk-reduces-inhaled-chlorine-triggered-ahr-and-airway-inflammation-in-the-mouse
#10
Yoichiro Hamamoto, Satoshi Ano, Benoit Allard, Michael O'Sullivan, Toby K McGovern, James G Martin
BACKGROUND AND PURPOSE: Cysteinyl leukotrienes (CysLTs) are pro-inflammatory lipid mediators that exacerbate disease state in several asthma phenotypes including asthma induced by allergen, virus and exercise. However, the role of CysLTs in irritant-induced airway disease is not well characterized. The purpose of the current study was to investigate the effect of montelukast (MK), a CysLT1R antagonist, on parameters of irritant-induced asthma (IIA) induced by inhalation of chlorine (Cl2 ) in the mouse...
July 18, 2017: British Journal of Pharmacology
https://www.readbyqxmd.com/read/28718842/secreted-protein-acidic-and-rich-in-cysteine-sparc-enhances-cell-proliferation-migration-and-epithelial-mesenchymal-transition-and-sparc-expression-is-associated-with-tumor-grade-in-head-and-neck-cancer
#11
Chih-Hau Chang, Meng-Chi Yen, Ssu-Hui Liao, Yu-Ling Hsu, Chung-Sheng Lai, Kao-Ping Chang, Ya-Ling Hsu
Secreted protein acidic and rich in cysteine (SPARC) is a secreted protein which is involved in various biological processes. SPARC expression is associated with tumor metastasis and poor prognosis in several types of cancer. However, the SPARC-induced signaling pathway was not fully understood in head and neck cancer. In this study, our results showed that SPARC treatment promoted cell proliferation and migration in head and neck cancer cell lines FaDu and Detroit 562. In addition, SPARC induced expression of epithelial mesenchymal transition (EMT) regulators, including Slug, Snail, and Twist in Detroit 562...
July 18, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28718412/stem-cells-cancer-and-musashi-in-blood-and-guts
#12
REVIEW
Michael G Kharas, Christopher J Lengner
The mammalian MSI family of RNA-binding proteins (RBPs) have important roles as oncoproteins in an array of tumor types, including leukemias, glioblastomas, and pancreatic, breast, lung, and colorectal cancers. The mammalian Msi genes, Msi1 and Msi2, have been most thoroughly investigated in two highly proliferative tissues prone to oncogenic transformation: the hematopoietic lineage and the intestinal epithelium. Despite their vast phenotypic differences, MSI proteins appear to have an analogous role in governing the stem cell compartment in both of these tissues, potentially providing a paradigm for a broader understanding of MSI function and oncogenic activities...
May 2017: Trends in Cancer
https://www.readbyqxmd.com/read/28718405/rna-binding-proteins-in-cancer-old-players-and-new-actors
#13
REVIEW
Bruno Pereira, Marc Billaud, Raquel Almeida
RNA-binding proteins (RBPs) are key players in post-transcriptional events. The combination of versatility of their RNA-binding domains with structural flexibility enables RBPs to control the metabolism of a large array of transcripts. Perturbations in RBP-RNA networks activity have been causally associated with cancer development, but the rational framework describing these contributions remains fragmented. We review here the evidence that RBPs modulate multiple cancer traits, emphasize their functional diversity, and assess future trends in the study of RBPs in cancer...
July 2017: Trends in Cancer
https://www.readbyqxmd.com/read/28717923/mirna-200a-induce-cell-apoptosis-in-renal-cell-carcinoma-by-directly-targeting-sirt1
#14
Hao Fu, Wenke Song, Xuancai Chen, Tao Guo, Bin Duan, Xinxi Wang, Yachun Tang, Liang Huang, Chi Zhang
Accumulating evidence indicates that microRNAs are implicated in tumor initiation and progression through negatively regulating oncogenes or tumor suppressor genes. In the present study, we report that the expression of miR-200a was significantly lower in renal cell carcinoma (RCC) specimens and RCC cell lines. Restoration of miR-200a suppressed cell growth, arrested cell cycle progression, and promoted cell apoptosis in RCC cell lines. We next used qRT-PCR array technology to identify Sirtuin 1 (SIRT1) as one of the downregulated proteins during miR-200a overexpression in 786-O cells...
July 17, 2017: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/28717582/in-vivo-multiphoton-imaging-of-a-diverse-array-of-fluorophores-to-investigate-deep-neurovascular-structure
#15
David R Miller, Ahmed M Hassan, Jeremy W Jarrett, Flor A Medina, Evan P Perillo, Kristen Hagan, S M Shams Kazmi, Taylor A Clark, Colin T Sullender, Theresa A Jones, Boris V Zemelman, Andrew K Dunn
We perform high-resolution, non-invasive, in vivo deep-tissue imaging of the mouse neocortex using multiphoton microscopy with a high repetition rate optical parametric amplifier laser source tunable between λ=1,100 and 1,400 nm. By combining the high repetition rate (511 kHz) and high pulse energy (400 nJ) of our amplifier laser system, we demonstrate imaging of vasculature labeled with Texas Red and Indocyanine Green, and neurons expressing tdTomato and yellow fluorescent protein. We measure the blood flow speed of a single capillary at a depth of 1...
July 1, 2017: Biomedical Optics Express
https://www.readbyqxmd.com/read/28716908/sequential-eviction-of-crowded-nucleoprotein-complexes-by-the-exonuclease-recbcd-molecular-motor
#16
Tsuyoshi Terakawa, Sy Redding, Timothy D Silverstein, Eric C Greene
In physiological settings, all nucleic acids motor proteins must travel along substrates that are crowded with other proteins. However, the physical basis for how motor proteins behave in these highly crowded environments remains unknown. Here, we use real-time single-molecule imaging to determine how the ATP-dependent translocase RecBCD travels along DNA occupied by tandem arrays of high-affinity DNA binding proteins. We show that RecBCD forces each protein into its nearest adjacent neighbor, causing rapid disruption of the protein-nucleic acid interaction...
July 17, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28716523/sensitivity-towards-the-grp78-inhibitor-kp1339-it-139-is-characterized-by-apoptosis-induction-via-caspase-8-upon-disruption-of-er-homeostasis
#17
Beatrix Schoenhacker-Alte, Thomas Mohr, Christine Pirker, Kushtrim Kryeziu, Paul-Steffen Kuhn, Alicia Buck, Thilo Hofmann, Christopher Gerner, Gerrit Hermann, Gunda Koellensperger, Bernhard K Keppler, Walter Berger, Petra Heffeter
The ruthenium drug and GRP78 inhibitor KP1339/IT-139 has already demonstrated promising anticancer activity in a phase I clinical trial. This study aimed to identify mechanisms underlying increased sensitivity to KP1339 treatment. Based on a screen utilizing 23 cell lines, a small panel was selected to compare KP1339-sensitive and low-responsive models. KP1339 sensitivity was neither based on differences in ruthenium accumulation, nor sensitivity to oxidative stress or constituents of KP1339 (ruthenium chloride and indazole)...
July 14, 2017: Cancer Letters
https://www.readbyqxmd.com/read/28715465/multiplexing-detection-of-igg-against-plasmodium-falciparum-pregnancy-specific-antigens
#18
Ana Maria Fonseca, Llorenç Quinto, Alfons Jiménez, Raquel González, Azucena Bardají, Sonia Maculuve, Carlota Dobaño, Maria Rupérez, Anifa Vala, John J Aponte, Esperanza Sevene, Eusebio Macete, Clara Menéndez, Alfredo Mayor
BACKGROUND: Pregnant women exposed to Plasmodium falciparum generate antibodies against VAR2CSA, the parasite protein that mediates adhesion of infected erythrocytes to the placenta. There is a need of high-throughput tools to determine the fine specificity of these antibodies that can be used to identify immune correlates of protection and exposure. Here we aimed at developing a multiplex-immunoassay to detect antibodies against VAR2CSA antigens. METHODS AND FINDINGS: We constructed two multiplex-bead arrays, one composed of 3 VAR2CSA recombinant-domains (DBL3X, DBL5Ɛ and DBL6Ɛ) and another composed of 46 new peptides covering VAR2CSA conserved and semi-conserved regions...
2017: PloS One
https://www.readbyqxmd.com/read/28715371/determining-cell-surface-expression-and-endocytic-rate-of-proteins-in-primary-astrocyte-cultures-using-biotinylation
#19
Daniel Kai Long Tham, Hakima Moukhles
Cell-surface proteins mediate a wide array of functions. In many cases, their activity is regulated by endocytic processes that modulate their levels at the plasma membrane. Here, we present detailed protocols for 2 methods that facilitate the study of such processes, both of which are based on the principle of the biotinylation of cell-surface proteins. The first is designed to allow for the semi-quantitative determination of the relative levels of a particular protein at the cell-surface. In it, the lysine residues of the plasma membrane proteins of cells are first labeled with a biotin moiety...
July 3, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28715156/neutrophil-mediated-enhancement-of-angiogenesis-and-osteogenesis-in-a-novel-triple-cell-co-culture-model-with-endothelial-cells-and-osteoblasts
#20
T D K Herath, Anis Larbi, Swee Hin Teoh, C James Kirkpatrick, Bee Tin Goh
Repair and regeneration of critical-sized bone defects remain a major challenge in orthopaedic and craniomaxillofacial surgery. Until now, attempts to bioengineer bone tissue have been hindered by the inability to establish proper angiogenesis and osteogenesis in the tissue construct. In the present study, we established a novel triple cell co-culture model consisting of osteoblasts, endothelial cells, and neutrophils and conducted a systematic investigation of the effects of neutrophils on angiogenesis and osteogenesis...
July 17, 2017: Journal of Tissue Engineering and Regenerative Medicine
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