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https://www.readbyqxmd.com/read/28654778/a-new-platform-for-serological-analysis-based-on-porous-3-dimensional-polyethylene-sinter-bodies
#1
Mohammed Alasel, Michael Keusgen
A new sensitive and selective platform, three-dimensional immunosensor, has been developed for a rapid serological diagnosis; detection of a Borrelia infection was considered as a model assay. The immunosensor is based on a 3-dimensional (3D) porous solid surface (sinter body) with dimensions of 2×2.5mm where a recombinant variable lipoprotein surface-exposed protein (VlsE; Borrelia-antigen) is immobilized by different techniques. The sinter body served as a robust and inexpensive carrier, which facilitated a successful hydrophobic adsorption as well as covalent immobilization of the antigen with sufficient amounts of on the surface...
June 20, 2017: Journal of Pharmaceutical and Biomedical Analysis
https://www.readbyqxmd.com/read/28654764/deep-proteome-profiling-reveals-common-prevalence-of-mzb1-positive-plasma-b-cells-in-human-lung-and-skin-fibrosis
#2
Herbert B Schiller, Christoph H Mayr, Gabriela Leuschner, Maximilian Strunz, Claudia Staab-Weijnitz, Stefan Preisendörfer, Beate Eckes, Pia Moinzadeh, Thomas Krieg, David A Schwartz, Rudolf A Hatz, Jürgen Behr, Matthias Mann, Oliver Eickelberg
RATIONALE: Analyzing the molecular heterogeneity of different forms of organ fibrosis may reveal common and specific factors and thus identify potential future therapeutic targets. OBJECTIVES: We sought to use proteome-wide profiling of human tissue fibrosis to (1) identify common and specific signatures across endstage interstitial lung disease (ILD) cases, (2) characterize ILD subgroups in an unbiased fashion, and (3) identify common and specific features of lung and skin fibrosis...
June 27, 2017: American Journal of Respiratory and Critical Care Medicine
https://www.readbyqxmd.com/read/28654654/identification-and-characterization-of-acka-dependent-protein-acetylation-in-neisseria-gonorrhoeae
#3
Deborah M B Post, Birgit Schilling, Lorri M Reinders, Alexandria K D'Souza, Margaret R Ketterer, Steven J Kiel, Aroon T Chande, Michael A Apicella, Bradford W Gibson
Neisseria gonorrhoeae, the causative agent of gonorrhea, has a number of factors known to contribute to pathogenesis; however, a full understanding of these processes and their regulation has proven to be elusive. Post-translational modifications (PTMs) of bacterial proteins are now recognized as one mechanism of protein regulation. In the present study, Western blot analyses, with an anti-acetyl-lysine antibody, indicated that a large number of gonococcal proteins are post-translationally modified. Previous work has shown that Nε-lysine acetylation can occur non-enzymatically with acetyl-phosphate (AcP) as the acetyl donor...
2017: PloS One
https://www.readbyqxmd.com/read/28654419/affimers-proteins-are-versatile-and-renewable-affinity-reagents
#4
Christian Tiede, Robert Bedford, Sophie J Heseltine, Gina Smith, Imeshi Wijetunga, Rebecca Ross, Danah AlQallaf, Ashley Pe Roberts, Alexander Balls, Alistair Curd, Ruth E Hughes, Heather Martin, Sarah R Needham, Laura C Zanetti-Domingues, Yashar Sadigh, Thomas P Peacock, Anna A Tang, Naomi Gibson, Hannah Kyle, Geoffrey W Platt, Nicola Ingram, Thomas Taylor, Louise P Coletta, Iain Manfield, Margaret Knowles, Sandra Bell, Filomena Esteves, Azhar Maqbool, Raj K Prasad, Mark Drinkhill, Robin S Bon, Vikesh Patel, Sarah A Goodchild, Marisa Martin-Fernandez, Ray J Owens, Joanne E Nettleship, Michael E Webb, Michael Harrison, Jonathan D Lippiat, Sreenivasan Ponnambalam, Michelle Peckham, Alastair Smith, Paul Ko Ferrigno, Matt Johnson, Michael J McPherson, Darren Charles Tomlinson
Molecular recognition reagents are key tools for understanding biological processes and are used universally by scientists to study protein expression, localisation and interactions. Antibodies remain the most widely used of such reagents and many show excellent performance, although some are poorly characterised or have stability or batch variability issues, supporting the use of alternative binding proteins as complementary reagents for many applications. Here we report on the use of Affimer proteins as research reagents...
June 27, 2017: ELife
https://www.readbyqxmd.com/read/28654217/dual-labelled-antibodies-for-flow-and-mass-cytometry-a-new-tool-for-cross-platform-comparison-and-enrichment-of-target-cells-for-mass-cytometry
#5
EDITORIAL
Sabine Baumgart, Axel R Schulz, Anette Peddinghaus, Silke Stanislawiak, Sarah Gillert, Heike Hirseland, Susanne Krauthäuser, Christian Dose, Henrik E Mei, Andreas Grützkau
Antibody conjugates applicable in both conventional flow and mass cytometry would offer interesting options for cross-platform comparison, as well as the enrichment of rare target cells by conventional flow cytometry (FC) sorting prior to deep phenotyping by mass cytometry (MC). Here, we introduce a simple method to generate dual fluorochrome/metal-labelled antibodies by consecutive orthogonal labelling. First we compared different fluorochrome-conjugated antibodies specific for CD4, such as FITC, Vio667, VioGreen or VioBlue for their compatibility with the conventional secondary MAXPAR(®) labelling protocol...
June 27, 2017: European Journal of Immunology
https://www.readbyqxmd.com/read/28654064/detection-and-visualization-of-dna-damage-induced-protein-complexes-in-suspension-cell-cultures-using-the-proximity-ligation-assay
#6
Mahnoush Bahjat, Timon A Bloedjes, Amélie van der Veen, Guus de Wilde, Chiel Maas, Jeroen E J Guikema
The DNA damage response orchestrates the repair of DNA lesions that occur spontaneously, are caused by genotoxic stress, or appear in the context of programmed DNA breaks in lymphocytes. The Ataxia-Telangiectasia Mutated kinase (ATM), ATM- and Rad3-Related kinase (ATR) and the catalytic subunit of DNA-dependent Protein Kinase (DNA-PKcs) are among the first that are activated upon induction of DNA damage, and are central regulators of a network that controls DNA repair, apoptosis and cell survival. As part of a tumor-suppressive pathway, ATM and ATR activate p53 through phosphorylation, thereby regulating the transcriptional activity of p53...
June 9, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28654062/using-a-whole-mount-immunohistochemical-method-to-study-the-innervation-of-the-biliary-tract-in-suncus-murinus
#7
Ke Ren, Yidan Dai, Kai Yi, Masanobu Kinoshita, Masahiro Itoh, Ichiro Sakata, Takafumi Sakai, Shuang-Qin Yi
This work describes the whole-mount immunohistochemistry staining method in detail, using neurofilament protein antibody to label the innervation of the biliary tract in Suncus murinus (S. murinus ). First, the specimen was dissected from S. murinus and fixed in 4% paraformaldehyde (PFA). Second, an enzymatic treatment and potential endogenous peroxidase inactivation were performed. The specimen was then exposed to the primary antibody, anti-neurofilament protein antibody, for 3-6 days. It was then incubated with the secondary antibody conjugated with horseradish peroxidase...
June 15, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28653853/post-translational-modifications-ptms-identified-on-endogenous-huntingtin-cluster-within-proteolytic-domains-between-heat-repeats
#8
Tamara Ratovitski, Robert N O'Meally, Mali Jiang, Raghothama Chaerkady, Ekaterine Chighladze, Jacqueline C Stewart, Xiaofang Wang, Nicolas Arbez, Elaine Roby, Athanasios Alexandris, Wenzhen Duan, Ravi Vijayvargia, Ihn Sik Seong, Daniel J Lavery, Robert N Cole, Christopher A Ross
Post-translational modifications (PTMs) of proteins regulate various cellular processes. PTMs of polyglutamine-expanded huntingtin (Htt) protein, which causes Huntington's disease (HD), are likely modulators of HD pathogenesis. Previous studies have identified and characterized several PTMs on exogenously expressed Htt fragments, but none of them were designed to systematically characterize PTMs on the endogenous full-length Htt protein. We found that full-length endogenous Htt, which was immunoprecipitated from HD knock-in mouse and human postmortem brain, is suitable for detection of PTMs by mass spectrometry...
June 27, 2017: Journal of Proteome Research
https://www.readbyqxmd.com/read/28653194/giardia-duodenalis-and-cryptosporidium-spp-as-contaminant-protozoa-of-the-main-rivers-of-western-romania-genetic-characterization-and-public-health-potential-of-the-isolates
#9
Kálmán Imre, Claudia Sala, Adriana Morar, Marius S Ilie, Judit Plutzer, Mirela Imre, Florin Ș Hora, Corina Badea, Mihai V Herbei, Gheorghe Dărăbuș
The objective of this study was to establish the prevalence, contamination level, and public health significance of Giardia duodenalis and Cryptosporidium spp. in the primary rivers of western Romania. A total of 53 sampling points in the 24 most important western Romanian rivers in four counties (Arad, Bihor, Caraș-Severin, and Timiș) were investigated from March to September 2016. Surface water samples were collected by microfiber filtration. Cryptosporidium and Giardia (oo)cysts were isolated using immunomagnetic separation (IMS) according to the USEPA 1623 method and, after staining with fluorescently labeled (FITC) monoclonal antibodies, were identified and counted under a microscope...
June 26, 2017: Environmental Science and Pollution Research International
https://www.readbyqxmd.com/read/28652982/reactivity-of-commercially-available-monoclonal-antibodies-to-human-cd-antigens-with-peripheral-blood-leucocytes-of-dromedary-camels-camelus-dromedarius
#10
Jamal Hussen, Turke Shawaf, Abdulkareem Imran Al-Herz, Hussain R Alturaifi, Ahmed M Alluwaimi
Monoclonal antibodies (mAbs) to cell surface molecules have been proven as a key tool for phenotypic and functional characterization of the cellular immune response. One of the major difficulties in studying camel cellular immunity consists in the lack of mAbs that dtect their leukocyte differentiation antigens. In the present study two-parameter flow cytometry was used to screen existing commercially available mAbs to human leukocyte antigens and major histocompatibility molecules (MHC) for their reactivity with camel leukocytes...
2017: Open veterinary journal
https://www.readbyqxmd.com/read/28652928/ph-of-anti-vegf-agents-in-the-human-vitreous-low-impact-of-very-different-formulations
#11
Bianka Sobolewska, Peter Heiduschka, Karl-Ulrich Bartz-Schmidt, Focke Ziemssen
BACKGROUND: The aim of the study was to measure pH changes of the human vitreous caused by the intravitreal drugs bevacizumab, ranibizumab, aflibercept, and ziv-aflibercept. METHODS: Fresh human vitreous samples were obtained during core vitrectomy (23-gauge) from patients with epiretinal gliosis. Aliquots of bevacizumab, ranibizumab, aflibercept or ziv-aflibercept (2 µl) were added consecutively to 200 µl of vitreous samples or 0.9% NaCl saline. The pH was measured using a pH-sensitive microelectrode...
2017: International Journal of Retina and Vitreous
https://www.readbyqxmd.com/read/28650478/a-dna-origami-platform-for-quantifying-protein-copy-number-in-super-resolution
#12
Francesca Cella Zanacchi, Carlo Manzo, Angel S Alvarez, Nathan D Derr, Maria F Garcia-Parajo, Melike Lakadamyali
Single-molecule-based super-resolution microscopy offers researchers a unique opportunity to quantify protein copy number with nanoscale resolution. However, while fluorescent proteins have been characterized for quantitative imaging using calibration standards, similar calibration tools for immunofluorescence with small organic fluorophores are lacking. Here we show that DNA origami, in combination with GFP antibodies, is a versatile platform for calibrating fluorophore and antibody labeling efficiency to quantify protein copy number in cellular contexts using super-resolution microscopy...
June 26, 2017: Nature Methods
https://www.readbyqxmd.com/read/28648630/resolution-of-a-steroid-resistant-hypereosinophilic-immune-diathesis-with-mepolizumab-and-concomitant-amelioration-of-a-mixed-thrombotic-microangiopathy
#13
J Justin Mulvey, Cynthia Magro, Amy Chadburn
The anaphylatoxins produced by an unbridled complement cascade in atypical hemolytic uremic syndrome (aHUS) can alter the leukocyte environment in tissues and peripheral blood, causing clinically significant eosinophilia. While the membrane attack complex and C5a anaphlatoxin can be suppressed with anti-C5 biologics, the production of C3a is still capable of driving a destructive hypereosinophilic syndrome in spite of anticomplement therapy. The side-effects of glucocorticoids in treating hypereosinophilic syndrome limit their therapeutic index in long-term treatment and this behooves the use of alternative strategies...
May 2, 2017: Blood Cells, Molecules & Diseases
https://www.readbyqxmd.com/read/28648614/the-effects-of-retinoic-acid-on-mmp-2-production-proliferation-and-ultrastructural-morphology-in-rat-uterus
#14
Tuğba Kotil, Şengül Şentürk, Türkan Sarioğlu, Leyla Tapul
AIMS: Retinoic acid (RA) has a vital importance in order to ensure continuity and morphology in many tissues. Matrix metalloproteinases (MMPs) have significant roles in proliferation, the formation of cancers, and metastasis. In this study the effects of RA on MMP-2 production in cells of rat uterus were investigated. METHODS: Twenty-four adult Spraque Dawley rats were divided into two groups, the experimental group was treated with 40mg/kg/day 13-cis RA for 5days by gavage...
June 22, 2017: Experimental and Toxicologic Pathology: Official Journal of the Gesellschaft Für Toxikologische Pathologie
https://www.readbyqxmd.com/read/28647167/adolescent-meningococcal-serogroup-a-w-and-y-immune-responses-following-immunization-with-quadrivalent-meningococcal-a-c-w-and-y-conjugate-vaccine-optimal-age-for-vaccination
#15
Mariëtte B van Ravenhorst, Fiona R M van der Klis, Debbie M van Rooijen, Elisabeth A M Sanders, Guy A M Berbers
BACKGROUND: Recently the incidence of meningococcal serogroup Y (MenY) and in particular serogroup W (MenW) invasive disease has risen in several European countries, including the Netherlands. Adolescents are a target group for primary prevention through vaccination to protect against disease and reduce carriage and induce herd protection in the population. The present study assessed MenA, MenW and MenY antibody levels in adolescents up to one year following primary vaccination with quadrivalent MenACWY-PS conjugated to tetanus toxoid (MenACWY-TT)...
June 21, 2017: Vaccine
https://www.readbyqxmd.com/read/28646725/eculizumab-decreases-the-procoagulant-activity-of-extracellular-vesicles-in-paroxysmal-nocturnal-hemoglobinuria-a-pilot-prospective-longitudinal-clinical-study
#16
Adeline Wannez, Bérangère Devalet, Céline Bouvy, Julie Laloy, Benoit Bihin, Bernard Chatelain, Christian Chatelain, Jean-Michel Dogné, François Mullier
INTRODUCTION: Paroxysmal nocturnal hemoglobinuria (PNH) is a disease characterized by the susceptibility of blood cells to attack by the complement system, inducing extracellular vesicle (EV) production. Thromboembolism is the leading cause of death in this condition. Eculizumab, a humanized monoclonal antibody which inhibits the C5 protein of the complement, reduces the thrombotic risk in PNH. MATERIALS AND METHOD: We conducted a pilot, prospective, open-label, longitudinal clinical study with six PNH patients treated with eculizumab...
June 9, 2017: Thrombosis Research
https://www.readbyqxmd.com/read/28645756/an-aptamer-based-pcr-method-coupled-with-magnetic-immunoseparation-for-sensitive-detection-of-salmonella-typhimurium-in-ground-turkey
#17
Lijun Wang, Ronghui Wang, Hong Wang, Michael Slavik, Hua Wei, Yanbin Li
Aptamers are single-stranded oligonucleotide ligands that can bind to targets with high affinity and specificity. They have been widely studied in the field of diagnostics as alternatives to antibodies due to their favorable features such as easy labeling, temperature tolerance, lower cost and recognition of a wide variety of targets. In this study, an aptamer-based PCR method coupled with magnetic immunoseparation was developed to detect S. Typhimurium from ground turkey. Firstly, biotinylated polyclonal anti-S...
June 20, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/28642244/investigation-of-protein-synthesis-in-drosophila-larvae-using-puromycin-labelling
#18
Lisa P Deliu, Abhishek Ghosh, Savraj S Grewal
Translational control of gene expression is an important regulator of growth, homeostasis and aging in Drosophila The ability to measure changes in protein synthesis in response to genetic and environmental cues is therefore important in studying these processes. Here we describe a simple and cost effective approach to assay protein synthesis in Drosophila larval cells and tissues. The method is based on the incorporation of puromycin into nascent peptide chains. Using an ex vivo approach, we label newly synthesized peptides in larvae with puromycin and then measure levels of new protein synthesis using an anti-puromycin antibody...
June 22, 2017: Biology Open
https://www.readbyqxmd.com/read/28641988/wavy-multistratified-amacrine-cells-in-the-monkey-retina-contain-immunoreactive-secretoneurin
#19
Andrea S Bordt, Ye Long, Nobuo Kouyama, Elizabeth S Yamada, Jens Hannibal, David W Marshak
The goals of this study were to describe the morphology, neurotransmitter content and synaptic connections of neurons in primate retinas that contain the neuropeptide secretoneurin. Amacrine cells were labeled with antibodies to secretoneurin in macaque and baboon retinas. Their processes formed three distinct plexuses in the inner plexiform layer: one in the outermost stratum, one in the center and one in the innermost stratum. In light microscopic double immunolabeling experiments, GABA was colocalized with secretoneurin in these cells, but glycine transporter 1 and Substance P were not...
June 19, 2017: Peptides
https://www.readbyqxmd.com/read/28640848/easy-labeling-of-proliferative-phase-and-sporogonic-phase-of-microsporidia-nosema-bombycis-in-host-cells
#20
Jie Chen, Wei Guo, Xiaoqun Dang, Yukang Huang, Fangyan Liu, Xianzhi Meng, Yaoyao An, Mengxian Long, Jialing Bao, Zeyang Zhou, Zhonghuai Xiang, Guoqing Pan
Microsporidia are eukaryotic, unicellular parasites that have been studied for more than 150 years. These organisms are extraordinary in their ability to invade a wide range of hosts including vertebrates and invertebrates, such as human and commercially important animals. A lack of appropriate labeling methods has limited the research of the cell cycle and protein locations in intracellular stages. In this report, an easy fluorescent labeling method has been developed to mark the proliferative and sporogonic phases of microsporidia Nosema bombycis in host cells...
2017: PloS One
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